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Sample records for die in-vivo messung

  1. In vivo characterization of microglial engulfment of dying neurons in the zebrafish spinal cord

    PubMed Central

    Morsch, Marco; Radford, Rowan; Lee, Albert; Don, Emily K.; Badrock, Andrew P.; Hall, Thomas E.; Cole, Nicholas J.; Chung, Roger

    2015-01-01

    Microglia are specialized phagocytes in the vertebrate central nervous system (CNS). As the resident immune cells of the CNS they play an important role in the removal of dying neurons during both development and in several neuronal pathologies. Microglia have been shown to prevent the diffusion of damaging degradation products of dying neurons by engulfment and ingestion. Here we describe a live imaging approach that uses UV laser ablation to selectively stress and kill spinal neurons and visualize the clearance of neuronal remnants by microglia in the zebrafish spinal cord. In vivo imaging confirmed the motile nature of microglia within the uninjured spinal cord. However, selective neuronal ablation triggered rapid activation of microglia, leading to phagocytic uptake of neuronal debris by microglia within 20–30 min. This process of microglial engulfment is highly dynamic, involving the extension of processes toward the lesion site and consequently the ingestion of the dying neuron. 3D rendering analysis of time-lapse recordings revealed the formation of phagosome-like structures in the activated microglia located at the site of neuronal ablation. This real-time representation of microglial phagocytosis in the living zebrafish spinal cord provides novel opportunities to study the mechanisms of microglia-mediated neuronal clearance. PMID:26379496

  2. Messung und Analyse

    NASA Astrophysics Data System (ADS)

    Bathelt, Hartmut; Scheinhardt, Michael; Sell, Hendrik; Sottek, Roland; Guidati, Sandro; Helfer, Martin

    Für die Beurteilung von Akustik und Fahrkomfort eines Fahrzeugs gilt in der Fahrzeugentwicklung immer noch der alte Grundsatz: "Der Kunde fährt nicht am Prüfstand, sondern auf der Straße“. Daher werden Gesamtbeurteilungen des Entwicklungsstandes und Konkurrenzvergleiche (Benchmarking) nach wie vor auf der Straße durchgeführt, meist auf ausgewählten Fahrbahnen am Prüfgelände oder im Rahmen der regelmäßigen Winter- und Sommererprobungen unter extremen Witterungsverhältnissen.

  3. Human NK cells proliferate and die in vivo more rapidly than T cells in healthy young and elderly adults.

    PubMed

    Lutz, Charles T; Karapetyan, Anush; Al-Attar, Ahmad; Shelton, Brent J; Holt, Kimberly J; Tucker, Jason H; Presnell, Steven R

    2011-04-15

    NK cells are essential for health, yet little is known about human NK turnover in vivo. In both young and elderly women, all NK subsets proliferated and died more rapidly than T cells. CD56(bright) NK cells proliferated rapidly but died relatively slowly, suggesting that proliferating CD56(bright) cells differentiate into CD56(dim) NK cells in vivo. The relationship between CD56(dim) and CD56(bright) proliferating cells indicates that proliferating CD56(dim) cells both self-renew and are derived from proliferating CD56(bright) NK cells. Our data suggest that some dying CD56(dim) cells become CD16(+)CD56(-) NK cells and that CD16(-)CD56(low) NK cells respond rapidly to cellular and cytokine stimulation. We propose a model in which all NK cell subsets are in dynamic flux. About half of CD56(dim) NK cells expressed CD57, which was weakly associated with low proliferation. Surprisingly, CD57 expression was associated with higher proliferation rates in both CD8(+) and CD8(-) T cells. Therefore, CD57 is not a reliable marker of senescent, nonproliferative T cells in vivo. NKG2A expression declined with age on both NK cells and T cells. Killer cell Ig-like receptor expression increased with age on T cells but not on NK cells. Although the percentage of CD56(bright) NK cells declined with age and the percentage of CD56(dim) NK cells increased with age, there were no significant age-related proliferation or apoptosis differences for these two populations or for total NK cells. In vivo human NK cell turnover is rapid in both young and elderly adults. PMID:21402893

  4. Loss of translation elongation factor (eEF1A2) expression in vivo differentiates between Wallerian degeneration and dying-back neuronal pathology

    PubMed Central

    Murray, Lyndsay M; Thomson, Derek; Conklin, Annalijn; Wishart, Thomas M; Gillingwater, Thomas H

    2008-01-01

    Wallerian degeneration and dying-back pathology are two well-known cellular pathways capable of regulating the breakdown and loss of axonal and synaptic compartments of neurons in vivo. However, the underlying mechanisms and molecular triggers of these pathways remain elusive. Here, we show that loss of translation elongation factor eEF1A2 expression in lower motor neurons and skeletal muscle fibres in homozygous Wasted mice triggered a dying-back neuropathy. Synaptic loss at the neuromuscular junction occurred in advance of axonal pathology and by a mechanism morphologically distinct from Wallerian degeneration. Dying-back pathology in Wasted mice was accompanied by reduced expression levels of the zinc finger protein ZPR1, as found in other dying-back neuropathies such as spinal muscular atrophy. Surprisingly, experimental nerve lesion revealed that Wallerian degeneration was significantly delayed in homozygous Wasted mice; morphological assessment revealed that ∼80% of neuromuscular junctions in deep lumbrical muscles at 24 h and ∼50% at 48 h had retained motor nerve terminals following tibial nerve lesion. This was in contrast to wild-type and heterozygous Wasted mice where < 5% of neuromuscular junctions had retained motor nerve terminals at 24 h post-lesion. These data show that eEF1A2 expression is required to prevent the initiation of dying-back pathology at the neuromuscular junction in vivo. In contrast, loss of eEF1A2 expression significantly inhibited the initiation and progression of Wallerian degeneration in vivo. We conclude that loss of eEF1A2 expression distinguishes mechanisms underlying dying-back pathology from those responsible for Wallerian degeneration in vivo and suggest that eEF1A2-dependent cascades may provide novel molecular targets to manipulate neurodegenerative pathways in lower motor neurons. PMID:19094180

  5. Das Neutron, der Kosmos und die Kräfte: Neutronen in der Teilchenphysik

    NASA Astrophysics Data System (ADS)

    Soldner, Torsten

    2003-05-01

    Das Neutron besitzt eine einzigartige Kombination von Eigenschaften. Sie ermöglicht die Untersuchung aller vier elementaren Kräfte. Dabei wurden beeindruckende Resultate erzielt, wie die Präzisionsmessungen der elektrischen Ladung des Neutrons oder der Feinstrukturkonstante zeigen. Die genaue Bestimmung der schwachen Wechselwirkungsstärke der Nukleonen liefert der Astrophysik wichtige Daten. Die Messung eines von Null verschiedenen elektrischen Dipolmoments des Neutrons könnte einen entscheidenden Hinweis über die Physik jenseits des Standardmodells der Teilchenphysik geben. Doch auch zur Aufklärung von Unsicherheiten innerhalb des Standardmodells selbst tragen Neutronen bei.

  6. Dying cells program their expedient disposal: serum amyloid P component upregulation in vivo and in vitro induced by photodynamic therapy of cancer.

    PubMed

    Merchant, Soroush; Sun, Jinghai; Korbelik, Mladen

    2007-12-01

    Serum amyloid P component (SAP) is known as a prototypic acute phase reactant in the mouse and the protein that binds to dying cells securing their swift disposal by phagocytes. Treatment of solid tumors by photodynamic therapy (PDT) triggers SAP production in the liver of host mice, its release in the circulation and accumulation in PDT-targeted lesions. In the present study, mouse Lewis lung carcinoma (LLC) cells treated in vitro by PDT are shown to upregulate their gene encoding SAP. This effect was manifested following PDT treatment mediated by various types of photosensitizers (Photofrin, BPD, mTHPC, ALA). Generated SAP protein was not detected in tissue supernatants but remained localized to producing PDT-treated cells. The upregulation of SAP gene was observed also in untreated IC-21 macrophages after they were co-incubated for 4 h with PDT-treated LLC cells. Based on these findings, SAP that accumulates in PDT-treated tumors may originate from both systemic sources (released from the liver as acute phase reactant) and local sources; the latter could include tumor cells directly sustaining PDT injury and macrophages invading the tumor that become stimulated by signals from these affected tumor cells. Since SAP gene upregulation in LLC cells increased with the lethality of PDT dose used for their treatment, we propose that cells sensing they are inflicted with mortal injury can turn on molecular programs insuring not only that they die an innocuous form of death (apoptosis) but also that once they are dead their elimination is (facilitated by SAP) swift and efficient. PMID:18046483

  7. SPHERICAL DIE

    DOEpatents

    Livingston, J.P.

    1959-01-27

    A die is presented for pressing powdered materials into a hemispherical shape of uniforin density and wall thickness comprising a fcmale and male die element held in a stationary spaced relation with the space being equivalent to the wall thickness and defining the hemispherical shape, a pressing ring linearly moveable along the male die element, an inlet to fill the space with powdered materials, a guiding system for moving the pressing ring along the male die element so as to press the powdered material and a heating system for heating the male element so that the powdered material is heated while being pressed.

  8. Desmosomes In Vivo

    PubMed Central

    Garrod, David

    2010-01-01

    The structure, function, and regulation of desmosomal adhesion in vivo are discussed. Most desmosomes in tissues exhibit calcium-independent adhesion, which is strongly adhesive or “hyperadhesive”. This is fundamental to tissue strength. Almost all studies in culture are done on weakly adhesive, calcium-dependent desmosomes, although hyperadhesion can be readily obtained in confluent cell culture. Calcium dependence is a default condition in vivo, found in wounds and embryonic development. Hyperadhesion appears to be associated with an ordered arrangement of the extracellular domains of the desmosomal cadherins, which gives rise to the intercellular midline identified in ultrastructural studies. This in turn probably depends on molecular order in the desmosomal plaque. Protein kinase C downregulates hyperadhesion and there is preliminary evidence that it may also be regulated by tyrosine kinases. Downregulation of desmosomes in vivo may occur by internalisation of whole desmosomes rather than disassembly. Hyperadhesion has implications for diseases such as pemphigus. PMID:20671997

  9. Nonimmune Chemotaxis In Vivo

    PubMed Central

    Wiener, Stanley; Lendvai, Sarai; Rogers, Brad; Urivetzky, Morton; Meilman, Edward

    1973-01-01

    Wistar rats were depleted of complement components using purified cobra venom factor (CoF) administered over a 30-hour period by intraperitoneal injection. Complement depletion was confirmed by immunodiffusion assay, hemolytic assay and inhibition of the reverse Arthus reaction. Rats depleted to below 3% of their normal serum complement level showed marked inhibition of chemotaxis into inflammatory fluid produced in polyvinyl sponges 24 hours after implantation into the dorsal subcutaneous region. Subsequent studies were done to test the effect of high local concentrations of CoF on the microcirculation, the neutrophil and the connective tissue. Local exposure of these tissues and cells to CoF in vivo had no inhibiting effect on chemotaxis. Cobra venom factor did not affect neutrophil survival in vitro or in vivo. The most likely hypothesis is that CoF works by depleting complement and that in vivo chemotaxis of the nonbacterial, nonimmune type is complement dependent to the extent of 60 to 80%. The system described allows for quantitative determination of chemotaxis in vivo. PMID:4767264

  10. EDITORIAL: Nanotechnology in vivo Nanotechnology in vivo

    NASA Astrophysics Data System (ADS)

    Demming, Anna

    2010-04-01

    -imaging labels [4]. A surface hydroxyl group renders silicon quantum dots soluble in water and the photoluminescence can be made stable with oxygen-passivation. In addition, researchers in Japan have demonstrated how the initially modest yield in the preparation of silicon quantum dots can be improved to tens of milligrams per batch, thus further promoting their application in bio-imaging [5]. In the search for non-toxic quantum dots, researchers at the Amrita Centre for Nanoscience in India have prepared heavy metal-free quantum dot bio-probes based on single phase ZnS [6]. The quantum dots are selectively doped with metals, transition metals and halides to provide tuneable luminescence properties, and they are surface conjugated with folic acid for cancer targeting. The quantum dots were demonstrated to be water-soluble, non-toxic in normal and cancer cell lines, and have bright, tuneable luminescence. So far most of the quantum dots developed for bio-imaging have had excitation and emission wavelengths in the visible spectrum, which is highly absorbed by tissue. This limits imaging with these quantum dots to superficial tissues. This week, researchers in China and the US reported work developing functionalized dots for in vivo tumour vasculature in the infrared part of the spectrum [7]. In addition the quantum dots were functionalised with glycine-aspartic acid (RGD) peptides, which target the vasculature of almost all types of growing tumours, unlike antibody- or aptamer-mediated targeting strategies that are specific to a particular cancer type. In this issue, researchers in China and the US demonstrate a novel type of contrast agent for ultrasonic tumour imaging [8]. Contrast-enhanced ultrasonic tumour imaging extends the diagnostic and imaging capabilities of traditional techniques. The use of nanoparticles as ultrasound contrast agents exploits the presence of open pores in the range of 380 to 780 nm in tumour blood vessels, which enhance the permeability and retention

  11. EDITORIAL: Nanotechnology in vivo Nanotechnology in vivo

    NASA Astrophysics Data System (ADS)

    Demming, Anna

    2010-04-01

    -imaging labels [4]. A surface hydroxyl group renders silicon quantum dots soluble in water and the photoluminescence can be made stable with oxygen-passivation. In addition, researchers in Japan have demonstrated how the initially modest yield in the preparation of silicon quantum dots can be improved to tens of milligrams per batch, thus further promoting their application in bio-imaging [5]. In the search for non-toxic quantum dots, researchers at the Amrita Centre for Nanoscience in India have prepared heavy metal-free quantum dot bio-probes based on single phase ZnS [6]. The quantum dots are selectively doped with metals, transition metals and halides to provide tuneable luminescence properties, and they are surface conjugated with folic acid for cancer targeting. The quantum dots were demonstrated to be water-soluble, non-toxic in normal and cancer cell lines, and have bright, tuneable luminescence. So far most of the quantum dots developed for bio-imaging have had excitation and emission wavelengths in the visible spectrum, which is highly absorbed by tissue. This limits imaging with these quantum dots to superficial tissues. This week, researchers in China and the US reported work developing functionalized dots for in vivo tumour vasculature in the infrared part of the spectrum [7]. In addition the quantum dots were functionalised with glycine-aspartic acid (RGD) peptides, which target the vasculature of almost all types of growing tumours, unlike antibody- or aptamer-mediated targeting strategies that are specific to a particular cancer type. In this issue, researchers in China and the US demonstrate a novel type of contrast agent for ultrasonic tumour imaging [8]. Contrast-enhanced ultrasonic tumour imaging extends the diagnostic and imaging capabilities of traditional techniques. The use of nanoparticles as ultrasound contrast agents exploits the presence of open pores in the range of 380 to 780 nm in tumour blood vessels, which enhance the permeability and retention

  12. In Vivo Antioxidant Assays.

    PubMed

    2016-01-01

    Oxidative stress and antioxidant deficiency have been implicated in the pathophysiology of a wide range of diseases and conditions. Consequently, over recent years many different supplementation trials have been implemented, aimed at improving clinical outcomes by boosting antioxidant levels. These trials included supplementation with individual antioxidants, antioxidant combinations, and antioxidant-rich foods such as fruit and vegetable juices and other plant extracts. To ensure that data from these trials are interpreted correctly, it is essential that suitable biomarkers are used to assess changes in in vivo antioxidant activity resulting from supplementation. Therefore, the measurement of antioxidant systems, such as superoxide dismutase, catalase, glutathione reductase, and status of other molecules in biological fluids with their quantification methods are simplified in this chapter. PMID:26939271

  13. In vivo dosimetry for IMRT

    SciTech Connect

    Vial, Philip

    2011-05-05

    In vivo dosimetry has a well established role in the quality assurance of 2D radiotherapy and 3D conformal radiotherapy. The role of in vivo dosimetry for IMRT is not as well established. IMRT introduces a range of technical issues that complicate in vivo dosimetry. The first decade or so of IMRT implementation has largely relied upon pre-treatment phantom based dose verification. During that time, several new devices and techniques for in vivo dosimetry have emerged with the promise of providing the ultimate form of IMRT dose verification. Solid state dosimeters continue to dominate the field of in vivo dosimetry in the IMRT era. In this report we review the literature on in vivo dosimetry for IMRT, with an emphasis on clinical evidence for different detector types. We describe the pros and cons of different detectors and techniques in the IMRT setting and the roles that they are likely to play in the future.

  14. Sputtered protective coatings for die casting dies

    NASA Technical Reports Server (NTRS)

    Mirtich, M. J.; Nieh, C. Y.; Wallace, J. F.

    1981-01-01

    This investigation determined whether selected ion beam sputtered coatings on H-13 die steel would have the potential of improving the thermal fatigue behavior of the steel used as a die in aluminum die casting. The coatings were selected to test candidate insulators and metals capable of providing protection of the die surface. The studies indicate that 1 micrometer thick W and Pt coatings reduced the thermal fatigue more than any other coating tested and are candidates to be used on a die surface to increase die life.

  15. Measuring In Vivo Mitophagy.

    PubMed

    Sun, Nuo; Yun, Jeanho; Liu, Jie; Malide, Daniela; Liu, Chengyu; Rovira, Ilsa I; Holmström, Kira M; Fergusson, Maria M; Yoo, Young Hyun; Combs, Christian A; Finkel, Toren

    2015-11-19

    Alterations in mitophagy have been increasingly linked to aging and age-related diseases. There are, however, no convenient methods to analyze mitophagy in vivo. Here, we describe a transgenic mouse model in which we expressed a mitochondrial-targeted form of the fluorescent reporter Keima (mt-Keima). Keima is a coral-derived protein that exhibits both pH-dependent excitation and resistance to lysosomal proteases. Comparison of a wide range of primary cells and tissues generated from the mt-Keima mouse revealed significant variations in basal mitophagy. In addition, we have employed the mt-Keima mice to analyze how mitophagy is altered by conditions including diet, oxygen availability, Huntingtin transgene expression, the absence of macroautophagy (ATG5 or ATG7 expression), an increase in mitochondrial mutational load, the presence of metastatic tumors, and normal aging. The ability to assess mitophagy under a host of varying environmental and genetic perturbations suggests that the mt-Keima mouse should be a valuable resource. PMID:26549682

  16. Nanocrystal targeting in vivo

    NASA Astrophysics Data System (ADS)

    Åkerman, Maria E.; Chan, Warren C. W.; Laakkonen, Pirjo; Bhatia, Sangeeta N.; Ruoslahti, Erkki

    2002-10-01

    Inorganic nanostructures that interface with biological systems have recently attracted widespread interest in biology and medicine. Nanoparticles are thought to have potential as novel intravascular probes for both diagnostic (e.g., imaging) and therapeutic purposes (e.g., drug delivery). Critical issues for successful nanoparticle delivery include the ability to target specific tissues and cell types and escape from the biological particulate filter known as the reticuloendothelial system. We set out to explore the feasibility of in vivo targeting by using semiconductor quantum dots (qdots). Qdots are small (<10 nm) inorganic nanocrystals that possess unique luminescent properties; their fluorescence emission is stable and tuned by varying the particle size or composition. We show that ZnS-capped CdSe qdots coated with a lung-targeting peptide accumulate in the lungs of mice after i.v. injection, whereas two other peptides specifically direct qdots to blood vessels or lymphatic vessels in tumors. We also show that adding polyethylene glycol to the qdot coating prevents nonselective accumulation of qdots in reticuloendothelial tissues. These results encourage the construction of more complex nanostructures with capabilities such as disease sensing and drug delivery.

  17. Wege in die Zukunft

    NASA Astrophysics Data System (ADS)

    Kauermann, Göran; Mosler, Karl

    Die Zukunft stellt große Herausforderungen an die Arbeit der Deutschen Statistischen Gesellschaft. Sie betreffen die gestiegenen Anforderungen der Nutzer von Statistik, die Kommunikationsmöglichkeiten des Internets sowie die Dynamik der statistischen Wissenschaften und ihrer Anwendungsgebiete. Das Kapitel 5 beschreibt, wie sich die Gesellschaft diesen Herausforderungen stellt und welche Ziele sie sich in der wissenschaftlichen Zusammenarbeit und im Kampf gegen das Innumeratentum gesetzt hat.

  18. Die drool and die drool theory

    NASA Astrophysics Data System (ADS)

    Schmalzer, A. M.; Giacomin, A. Jeffrey

    2013-04-01

    When molten plastic is extruded from a die, it sometimes collects on the open face of the die. Known as die drool, this phenomenon costs plastics manufacturers by requiring die cleaning. This has been attributed to many causes, but none of these has led to an equation for the drool rate. In this work we provide an exact analytical solution for the drool rate, and we base this solution on a postulate of a cohesive slip layer near the die walls. We thus attribute die drool to cohesive failure within the fluid at an internal surface where the fluid slips on itself. We adimensionalize the drool rate with the production rate, and call this the build up ratio, BR. We provide an exact analytical solution for BR when the cohesive slip layer either sticks at the wall. We examine the slit geometry corresponding to sheet or film extrusion.

  19. Displaced capillary dies

    DOEpatents

    Kalejs, Juris P.; Chalmers, Bruce; Surek, Thomas

    1984-01-01

    An asymmetrical shaped capillary die made exclusively of graphite is used to grow silicon ribbon which is capable of being made into solar cells that are more efficient than cells produced from ribbon made using a symmetrically shaped die.

  20. Displaced capillary dies

    DOEpatents

    Kalejs, Juris P.; Chalmers, Bruce; Surek, Thomas

    1982-01-01

    An asymmetrical shaped capillary die made exclusively of graphite is used to grow silicon ribbon which is capable of being made into solar cells that are more efficient than cells produced from ribbon made using a symmetrically shaped die.

  1. Packaged die heater

    SciTech Connect

    Spielberger, Richard; Ohme, Bruce Walker; Jensen, Ronald J.

    2011-06-21

    A heater for heating packaged die for burn-in and heat testing is described. The heater may be a ceramic-type heater with a metal filament. The heater may be incorporated into the integrated circuit package as an additional ceramic layer of the package, or may be an external heater placed in contact with the package to heat the die. Many different types of integrated circuit packages may be accommodated. The method provides increased energy efficiency for heating the die while reducing temperature stresses on testing equipment. The method allows the use of multiple heaters to heat die to different temperatures. Faulty die may be heated to weaken die attach material to facilitate removal of the die. The heater filament or a separate temperature thermistor located in the package may be used to accurately measure die temperature.

  2. Experimental Cryosurgery Investigations In Vivo

    PubMed Central

    Gage, A.A.; Baust, J.M.; Baust, J.G.

    2009-01-01

    Cryosurgery is the use of freezing temperatures to elicit an ablative response in a targeted tissue. This review provides a global overview of experimentation in vivo which has been the basis of advancement of this widely applied therapeutic option. The cellular and tissue-related events that underlie the mechanisms of destruction, including direct cell injury (cryolysis), vascular stasis, apoptosis and necrosis, are described and are related to the optimal methods of technique of freezing to achieve efficacious therapy. In vivo experiments with major organs, including wound healing, the putative immunological response following thawing, and the use of cryoadjunctive strategies to enhance cancer cell sensitivity to freezing, are described. PMID:19833119

  3. HIGH PRESSURE DIES

    DOEpatents

    Wilson, W.B.

    1960-05-31

    A press was invented for subjecting specimens of bismuth, urania, yttria, or thoria to high pressures and temperatures. The press comprises die parts enclosing a space in which is placed an electric heater thermally insulated from the die parts so as not to damage them by heat. The die parts comprise two opposed inner frustoconical parts and an outer part having a double frustoconical recess receiving the inner parts. The die space decreases in size as the inner die parts move toward one another against the outer part and the inner parts, though very hard, do not fracture because of the mode of support provided by the outer part.

  4. The In Vivo Salmonella gallinarum

    PubMed Central

    Buxton, A.

    1959-01-01

    The intravenous inoculation into fowls of large doses of crude polysaccharide preparations derived from a smooth strain of Salmonella gallinarum resulted in the adsorption of some of the polysaccharide on the surfaces of circulating erythrocytes. This in vivo adsorption, detectable by an antiglobulin haemagglutination test, lasted for a maximum of approximately 24 hours after inoculation of polysaccharide. Similarly, in vivo sensitization of avian erythrocytes reached a maximum at about 8-10 days following oral infection of fowls with Salm. gallinarum. These results are discussed in relation to the ability of the reticulo-endothelial system to remove polysaccharide from the blood stream, and to the possible consequences of the hosts' immunological response to sensitized erythrocytes. PMID:13806563

  5. Terahertz pulsed imaging in vivo

    NASA Astrophysics Data System (ADS)

    Pickwell-MacPherson, E.

    2011-03-01

    Terahertz (1012 Hz) pulsed imaging is a totally non-destructive and non-ionising imaging modality and thus potential applications in medicine are being investigated. In this paper we present results using our hand-held terahertz probe that has been designed for in vivo use. In particular, we use the terahertz probe to perform reflection geometry in vivo measurements of human skin. The hand-held terahertz probe gives more flexibility than a typical flat-bed imaging system, but it also results in noisier data and requires existing processing methods to be improved. We describe the requirements and limitations of system geometry, data acquisition rate, image resolution and penetration depth and explain how various factors are dependent on each other. We show how some of the physical limitations can be overcome using novel data processing methods.

  6. In vivo imaging of sulfotransferases

    DOEpatents

    Barrio, Jorge R; Kepe, Vladimir; Small, Gary W; Satyamurthy, Nagichettiar

    2013-02-12

    Radiolabeled tracers for sulfotransferases (SULTs), their synthesis, and their use are provided. Included are substituted phenols, naphthols, coumarins, and flavones radiolabeled with .sup.18F, .sup.123I, .sup.124I, .sup.125I, or .sup.11C. Also provided are in vivo techniques for using these and other tracers as analytical and diagnostic tools to study sulfotransferase distribution and activity, in health and disease, and to evaluate therapeutic interventions.

  7. Die singulation method

    SciTech Connect

    Swiler, Thomas P; Garcia, Ernest J; Francis, Kathryn M

    2014-01-07

    A method is disclosed for singulating die from a semiconductor substrate (e.g. a semiconductor-on-insulator substrate or a bulk silicon substrate) containing an oxide layer (e.g. silicon dioxide or a silicate glass) and one or more semiconductor layers (e.g. monocrystalline or polycrystalline silicon) located above the oxide layer. The method etches trenches through the substrate and through each semiconductor layer about the die being singulated, with the trenches being offset from each other around at least a part of the die so that the oxide layer between the trenches holds the substrate and die together. The trenches can be anisotropically etched using a Deep Reactive Ion Etching (DRIE) process. After the trenches are etched, the oxide layer between the trenches can be etched away with a HF etchant to singulate the die. A release fixture can be located near one side of the substrate to receive the singulated die.

  8. Die singulation method

    SciTech Connect

    Swiler, Thomas P.; Garcia, Ernest J.; Francis, Kathryn M.

    2013-06-11

    A method is disclosed for singulating die from a semiconductor substrate (e.g. a semiconductor-on-insulator substrate or a bulk silicon substrate) containing an oxide layer (e.g. silicon dioxide or a silicate glass) and one or more semiconductor layers (e.g. monocrystalline or polycrystalline silicon) located above the oxide layer. The method etches trenches through the substrate and through each semiconductor layer about the die being singulated, with the trenches being offset from each other around at least a part of the die so that the oxide layer between the trenches holds the substrate and die together. The trenches can be anisotropically etched using a Deep Reactive Ion Etching (DRIE) process. After the trenches are etched, the oxide layer between the trenches can be etched away with an HF etchant to singulate the die. A release fixture can be located near one side of the substrate to receive the singulated die.

  9. Extrusion die and method

    DOEpatents

    Lipp, G. Daniel

    1994-04-26

    A method and die apparatus for manufacturing a honeycomb body of rhombic cell cross-section by extrusion through an extrusion die of triangular cell discharge slot configuration, the die incorporating feedholes at selected slot intersections only, such that slot segments communicating directly with the feedholes discharge web material and slot segments not so connected do not discharge web material, whereby a rhombic cell cross-section in the extruded body is provided.

  10. The Ambiguous Dying Syndrome

    ERIC Educational Resources Information Center

    Bern-Klug, Mercedes

    2004-01-01

    More than one-half of the 2.4 million deaths that will occur in the United States in 2004 will be immediately preceded by a time in which the likelihood of dying can best be described as "ambiguous." Many people die without ever being considered "dying" or "at the end of life." These people may miss out on the opportunity to close important…

  11. Sputtered protective coatings for die casting dies

    NASA Technical Reports Server (NTRS)

    Mirtich, M. J.; Nieh, C.-Y.; Wallace, J. F.

    1981-01-01

    Three experimental research designs investigating candidate materials and processes involved in protective die surface coating procedures by sputter deposition, using ion beam technologies, are discussed. Various pre-test results show that none of the coatings remained completely intact for 15,000 test cycles. The longest lifetime was observed for coatings such as tungsten, platinum, and molybdenum which reduced thermal fatigue, but exhibited oxidation and suppressed crack initiation only as long as the coating did not fracture. Final test results confirmed earlier findings and coatings with Pt and W proved to be the candidate materials to be used on a die surface to increase die life. In the W-coated specimens, which remained intact on the surface after thermal fatigue testing, no oxidation was found under the coating, although a few cracks formed on the surface where the coating broke down. Further research is planned.

  12. Die Soldering in Aluminium Die Casting

    SciTech Connect

    Han, Q.; Kenik, E.A.; Viswanathan, S.

    2000-03-15

    Two types of tests, dipping tests and dip-coating tests were carried out on small steel cylinders using pure aluminum and 380 alloy to investigate the mechanism of die soldering during aluminum die casting. Optical and scanning electron microscopy were used to study the morphology and composition of the phases formed during soldering. A soldering mechanism is postulated based on experimental observations. A soldering critical temperature is postulated at which iron begins to react with aluminum to form an aluminum-rich liquid phase and solid intermetallic compounds. When the temperature at the die surface is higher than this critical temperature, the aluminum-rich phase is liquid and joins the die with the casting during the subsequent solidification. The paper discusses the mechanism of soldering for the case of pure aluminum and 380 alloy casting in a steel mold, the factors that promote soldering, and the strength of the bond formed when soldering occurs. conditions, an aluminum-rich soldering layer may also form over the intermetallic layer. Although a significant amount of research has been conducted on the nature of these intermetallics, little is known about the conditions under which soldering occurs.

  13. Is Dying Young Worse than Dying Old?

    ERIC Educational Resources Information Center

    Jecker, Nancy S.; Schneiderman, Lawrence J.

    1994-01-01

    Notes that, in contemporary Western society, people feel death of small child is greater injustice than death of older adult and experience correspondingly greater sorrow, anger, regret, or bitterness when very young person dies. Contrasts these attitudes with those of ancient Greece and shows relevance that different attitudes toward death have…

  14. Extrusion die and method

    DOEpatents

    Lipp, G. Daniel

    1994-05-03

    A method and die apparatus for manufacturing a honeycomb body of triangular cell cross-section and high cell density, the die having a combination of (i) feedholes feeding slot intersections and (ii) feedholes feeding slot segments not supplied from slot intersections, whereby a reduction in feedhole count is achieved while still retaining good extrusion efficiency and extrudate uniformity.

  15. Micromechanical die attachment surcharge

    DOEpatents

    Filter, William F.; Hohimer, John P.

    2002-01-01

    An attachment structure is disclosed for attaching a die to a supporting substrate without the use of adhesives or solder. The attachment structure, which can be formed by micromachining, functions purely mechanically in utilizing a plurality of shaped pillars (e.g. round, square or polygonal and solid, hollow or slotted) that are formed on one of the die or supporting substrate and which can be urged into contact with various types of mating structures including other pillars, a deformable layer or a plurality of receptacles that are formed on the other of the die or supporting substrate, thereby forming a friction bond that holds the die to the supporting substrate. The attachment structure can further include an alignment structure for precise positioning of the die and supporting substrate to facilitate mounting the die to the supporting substrate. The attachment structure has applications for mounting semiconductor die containing a microelectromechanical (MEM) device, a microsensor or an integrated circuit (IC), and can be used to form a multichip module. The attachment structure is particularly useful for mounting die containing released MEM devices since these devices are fragile and can otherwise be damaged or degraded by adhesive or solder mounting.

  16. Colchicine prevents tumor necrosis factor-induced toxicity in vivo.

    PubMed Central

    Tiegs, G; Freudenberg, M A; Galanos, C; Wendel, A

    1992-01-01

    Tumor necrosis factor (TNF) toxicity was induced in vivo by intravenous administration of 15 micrograms of recombinant murine TNF-alpha per kg to galactosamine-sensitized mice. Within 8 h, the animals developed a fulminant hepatitis. Intravenous administration of 0.5 mg of colchicine per kg at 19 and 4 h prior to TNF challenge protected the animals against hepatitis. Lipopolysaccharide (LPS)-stimulated, bone marrow-derived macrophages from C3H/HeN mice released significant amounts of TNF in vitro. When such macrophages were intravenously given to LPS-resistant galactosamine-sensitized C3H/HeJ mice, these animals died within 24 h. Preincubation of these transferred macrophages with colchicine did not suppress the LPS-inducible TNF release from these cells. Concordantly, administration of macrophages exposed to colchicine in vitro resulted in full lethality. However, in vivo pretreatment of C3H/HeJ mice with colchicine 19 and 4 h prior to the transfer of LPS-stimulated macrophages prevented lethality. In LPS-responsive NMRI mice which had been protected against galactosamine-LPS-induced hepatitis by pretreatment with colchicine, TNF was still released into the blood. We conclude from our findings that the in vivo protection by colchicine is mediated by blocking TNF action on target cells while the effector cells of LPS toxicity, i.e., the macrophages, remain responsive. PMID:1563785

  17. Towards a disposable in vivo miniature implantable fluorescence detector

    NASA Astrophysics Data System (ADS)

    Bellis, Stephen; Jackson, J. Carlton; Mathewson, Alan

    2006-02-01

    In the field of fluorescent microscopy, neuronal activity, diabetes and drug treatment are a few of the wide ranging biomedical applications that can be monitored with the use of dye markers. Historically, in-vivo fluorescent detectors consist of implantable probes coupled by optical fibre to sophisticated bench-top instrumentation. These systems typically use laser light to excite the fluorescent marker dies and using sensors, such as the photo-multiplier tube (PMT) or charge coupled devices (CCD), detect the fluorescent light that is filtered from the total excitation. Such systems are large and expensive. In this paper we highlight the first steps toward a fully implantable in-vivo fluorescence detection system. The aim is to make the detector system small, low cost and disposable. The current prototype is a hybrid platform consisting of a vertical cavity surface emitting laser (VCSEL) to provide the excitation and a filtered solid state Geiger mode avalanche photo-diode (APD) to detect the emitted fluorescence. Fluorescence detection requires measurement of extremely low levels of light so the proposed APD detectors combine the ability to count individual photons with the added advantage of being small in size. At present the exciter and sensor are mounted on a hybrid PCB inside a 3mm diameter glass tube.This is wired to external electronics, which provide quenching, photon counting and a PC interface. In this configuration, the set-up can be used for in-vitro experimentation and in-vivo analysis conducted on animals such as mice.

  18. Hydroxyl radical detection in vivo

    SciTech Connect

    Chevion, M.; Floyd, R.A.

    1986-05-01

    Hydroxyl radicals have been implicated as the actual species responsible for the deleterious effects of active oxygen in biology. However, in most cases, its presence has only been inferred by circumstantial evidence. Using electrochemical detection coupled to HPLC separation technique the authors can identify and quantitate (at sub-picomole level) the hydroxylated products of 3 aromatic compounds (phenol, salicylate, and 2-deoxy-guanosine) as a direct measure of hydroxyl radical formation. Firstly, the authors showed that mixing ascorbate with copper ions (in the absence of presence of a protein) yields catechols, dihydroxybenzoic acids and 8-OH-deoxy-guanosine (8-OHdG). This approach has been used to study the formation of OH in vivo. Human granulocytes stimulated with TPA showed that 8-OHdG was formed in the cellular DNA at high levels (one 8-OHdG/800 DNA bases). Unstimulated granulocytes contained 8-OHdG below detection level. Formation of 8-OHdG in the TPA-stimulated granulocytes DNA was decreased by the addition of SOD and catalase. Using salicylate as an in vivo scavenger of hydroxyl radicals the authors showed that the level of trapped-dihydroxybenzoic acids is increased approx.8 and approx.3 fold in the lungs and liver of paraquat-poisoned mice, respectively, as compared to normal animals. Similarly, the detected level of dihydroxybenzoic acids in the hearts of adriamycin-treated rats was increased over 100-fold as compared to the hearts of control animals.

  19. Papillomavirus DNA Complementation in Vivo

    PubMed Central

    Hu, Jiafen; Cladel, Nancy M.; Budgeon, Lynn; Balogh, Karla K.; Christensen, Neil D.

    2009-01-01

    Recent phylogenic studies indicate that DNA recombination could have occurred in ancient papillomaviruses types. However, no experimental data is available to demonstrate this event because of the lack of human papillomavirus infection models. We have used the cottontail rabbit papillomavirus (CRPV)/rabbit model to study pathogenesis and immunogenicity of different mutant genomes in vivo. Although the domestic rabbit is not a natural host for CRPV infection, it is possible to initiate infection with naked CRPV DNA cloned into a plasmid and monitor papilloma outgrowth on these animals. Taking advantage of a large panel of mutants based on a CRPV strain (Hershey CRPV), we tested the hypothesis that two non-viable mutant genomes could induce papillomas by either recombination or complementation. We found that co-infection with a dysfunctional mutant with an E2 transactivation domain mutation and another mutant with an E7 ATG knock out generated papillomas in rabbits. DNA extracted from these papillomas contained genotypes from both parental genomes. Three additional pairs of dysfunctional mutants also showed similar results. Individual wild type genes were also shown to rescue the function of corresponding dysfunctional mutants. Therefore, we suggest that complementation occurred between these two non-viable mutant PV genomes in vivo. PMID:19379784

  20. Die zwei Kulturen

    NASA Astrophysics Data System (ADS)

    Ankolekar, Anupriya; Krötzsch, Markus; Tran, Than; Vrandecic, Denny

    Oft werden zwei mögliche Entwicklungen des Webs diskutiert - das Web 2.0 und das Semantic Web. Wenn wir diese zwei Visionen für das zukünftige Web unter die Lupe nehmen, dann lässt sich feststellen, dass sich die Ideen in ihrem Kern und ihren Technologien gegenseitig ergänzen. Dementsprechend können und sollen beide Visionen von den Erfahrungen und Stärken der anderen profitieren. Wir glauben daran, dass zukünftige Webanwendungen den Web 2.0-Fokus auf Community und Benutzerfreundlichkeit beibehalten und, darüber hinaus, auch von Technologien des Semantic Web zur Vereinfachung der mashupähnlichen Datenintegration profitieren werden. Auf Basis eines Semantic Blog-Szenarios werden wir hier die Vorteile einer möglichen Kombination von Semantic Web und Web 2.0 illustrieren, die zeitnah realisiert werden kann. Wir werden auch auf technische Probleme eingehen, die bei der Erweiterung dieses Szenarios entstehen. Wir stellen dar, wie aktuelle Entwicklungen in der Semantic Web Forschung diese Probleme angehen können, und setzen zugleich auch Schwerpunkte für die zukünftige Forschung, die in diesem Zusammenhang relevant sind.

  1. Measuring Vascular Permeability In Vivo.

    PubMed

    Meijer, Eelco F J; Baish, James W; Padera, Timothy P; Fukumura, Dai

    2016-01-01

    Over the past decades, in vivo vascular permeability measurements have provided significant insight into vascular functions in physiological and pathophysiological conditions such as the response to pro- and anti-angiogenic signaling, abnormality of tumor vasculature and its normalization, and delivery and efficacy of therapeutic agents. Different approaches for vascular permeability measurements have been established. Here, we describe and discuss a conventional 2D imaging method to measure vascular permeability, which was originally documented by Gerlowski and Jain in 1986 (Microvasc Res 31:288-305, 1986) and further developed by Yuan et al. in the early 1990s (Microvasc Res 45:269-289, 1993; Cancer Res 54:352-3356, 1994), and our recently developed 3D imaging method, which advances the approach originally described by Brown et al. in 2001 (Nat Med 7:864-868, 2001). PMID:27581015

  2. [Towards optical in vivo electrophysiology].

    PubMed

    Lambot, Laurie; Gall, David

    Optical imaging of voltage indicators is a promising approach for detecting the activity of neuronal circuits with high spatial and temporal resolution. In this context, genetically encoded voltage indicators, combining genetic targeting and optical readout of transmembrane voltage, represent a technological breaktrough that will without doubt have a major impact in neuroscience. However, so far the existing genetically encoded voltage indicators lacked the capabilities to detect individual action potentials and fast spike trains in live animals. Here, we present a novel indicator allowing high-fidelity imaging of individual spikes and dentritic voltage dynamics in vivo. Used in combination with optogenetics, which allows to manipulate neuronal activity, this opens the possibility of an all-optical electrophysiology. PMID:27615186

  3. In Vivo EPR For Dosimetry

    PubMed Central

    Swartz, Harold M.; Burke, Greg; Coey, M.; Demidenko, Eugene; Dong, Ruhong; Grinberg, Oleg; Hilton, James; Iwasaki, Akinori; Lesniewski, Piotr; Kmiec, Maciej; Lo, Kai-Ming; Nicolalde, R. Javier; Ruuge, Andres; Sakata, Yasuko; Sucheta, Artur; Walczak, Tadeusz; Williams, Benjamin B.; Mitchell, Chad; Romanyukha, Alex; Schauer, David A.

    2007-01-01

    As a result of terrorism, accident, or war, populations potentially can be exposed to doses of ionizing radiation that could cause direct clinical effects within days or weeks. There is a critical need to determine the magnitude of the exposure to individuals so that those with significant risk have appropriate procedures initiated immediately, while those without a significant probability of acute effects can be reassured and removed from the need for further consideration in the medical/emergency system. In many of the plausible scenarios there is an urgent need to make the determination very soon after the event and while the subject is still present. In vivo EPR measurements of radiation-induced changes in the enamel of teeth is a method, perhaps the only such method, which can differentiate among doses sufficiently for classifying individuals into categories for treatment with sufficient accuracy to facilitate decisions on medical treatment. In its current state, the in vivo EPR dosimeter can provide estimates of absorbed dose with an error approximately ± 50 cGy over the range of interest for acute biological effects of radiation, assuming repeated measurements of the tooth in the mouth of the subject. The time required for acquisition, the lower limit, and the precision are expected to improve, with improvements in the resonator and the algorithm for acquiring and calculating the dose. The magnet system that is currently used, while potentially deployable, is somewhat large and heavy, requiring that it be mounted on a small truck or trailer. Several smaller magnets, including an intraoral magnet are under development, which would extend the ease of use of this technique. PMID:18591988

  4. Die Zeitung der Zukunft

    NASA Astrophysics Data System (ADS)

    Wieser, Christoph; Schaffert, Sebastian

    Schon lange wird spekuliert, wie wir in Zukunft Zeitung lesen werden. Werden wir am Frühstückstisch wie gewohnt in einer Zeitung aus Papier schmökern oder werden wir die Zeitung als biegsame Folie beschrieben mit elektronischer Tinte in Händen halten? Wird die Zeitung mit anderen Medien wie Radio und Fernsehen verschmelzen? Viele Varianten sind denkbar. Heute lässt sich schon ein Trend ablesen: Immer mehr Leser entdecken die Online-Zeitung als Informationsmedium, eine Voraussetzung für die Nutzung neuer Technologien in der Zeitung der Zukunft. In diesem Kapitel stellen wir Entwicklungsmöglichkeiten der Online-Zeitung dar, wie sie im Social Semantic Web möglich werden.

  5. When Somebody Dies

    MedlinePlus

    ... alguien muere All living things — including bugs and fish and people — die. It's difficult, even for grownups, ... kind of death for families and friends to deal with because it happens so fast. There is ...

  6. Coffee induces autophagy in vivo

    PubMed Central

    Pietrocola, Federico; Malik, Shoaib Ahmad; Mariño, Guillermo; Vacchelli, Erika; Senovilla, Laura; Chaba, Kariman; Niso-Santano, Mireia; Maiuri, Maria Chiara; Madeo, Frank; Kroemer, Guido

    2014-01-01

    Epidemiological studies and clinical trials revealed that chronic consumption coffee is associated with the inhibition of several metabolic diseases as well as reduction in overall and cause-specific mortality. We show that both natural and decaffeinated brands of coffee similarly rapidly trigger autophagy in mice. One to 4 h after coffee consumption, we observed an increase in autophagic flux in all investigated organs (liver, muscle, heart) in vivo, as indicated by the increased lipidation of LC3B and the reduction of the abundance of the autophagic substrate sequestosome 1 (p62/SQSTM1). These changes were accompanied by the inhibition of the enzymatic activity of mammalian target of rapamycin complex 1 (mTORC1), leading to the reduced phosphorylation of p70S6K, as well as by the global deacetylation of cellular proteins detectable by immunoblot. Immunohistochemical analyses of transgenic mice expressing a GFP–LC3B fusion protein confirmed the coffee-induced relocation of LC3B to autophagosomes, as well as general protein deacetylation. Altogether, these results indicate that coffee triggers 2 phenomena that are also induced by nutrient depletion, namely a reduction of protein acetylation coupled to an increase in autophagy. We speculate that polyphenols contained in coffee promote health by stimulating autophagy. PMID:24769862

  7. In Vivo Facilitated Diffusion Model

    PubMed Central

    Bauer, Maximilian; Metzler, Ralf

    2013-01-01

    Under dilute in vitro conditions transcription factors rapidly locate their target sequence on DNA by using the facilitated diffusion mechanism. However, whether this strategy of alternating between three-dimensional bulk diffusion and one-dimensional sliding along the DNA contour is still beneficial in the crowded interior of cells is highly disputed. Here we use a simple model for the bacterial genome inside the cell and present a semi-analytical model for the in vivo target search of transcription factors within the facilitated diffusion framework. Without having to resort to extensive simulations we determine the mean search time of a lac repressor in a living E. coli cell by including parameters deduced from experimental measurements. The results agree very well with experimental findings, and thus the facilitated diffusion picture emerges as a quantitative approach to gene regulation in living bacteria cells. Furthermore we see that the search time is not very sensitive to the parameters characterizing the DNA configuration and that the cell seems to operate very close to optimal conditions for target localization. Local searches as implied by the colocalization mechanism are only found to mildly accelerate the mean search time within our model. PMID:23349772

  8. Coffee induces autophagy in vivo.

    PubMed

    Pietrocola, Federico; Malik, Shoaib Ahmad; Mariño, Guillermo; Vacchelli, Erika; Senovilla, Laura; Chaba, Kariman; Niso-Santano, Mireia; Maiuri, Maria Chiara; Madeo, Frank; Kroemer, Guido

    2014-01-01

    Epidemiological studies and clinical trials revealed that chronic consumption coffee is associated with the inhibition of several metabolic diseases as well as reduction in overall and cause-specific mortality. We show that both natural and decaffeinated brands of coffee similarly rapidly trigger autophagy in mice. One to 4 h after coffee consumption, we observed an increase in autophagic flux in all investigated organs (liver, muscle, heart) in vivo, as indicated by the increased lipidation of LC3B and the reduction of the abundance of the autophagic substrate sequestosome 1 (p62/SQSTM1). These changes were accompanied by the inhibition of the enzymatic activity of mammalian target of rapamycin complex 1 (mTORC1), leading to the reduced phosphorylation of p70(S6K), as well as by the global deacetylation of cellular proteins detectable by immunoblot. Immunohistochemical analyses of transgenic mice expressing a GFP-LC3B fusion protein confirmed the coffee-induced relocation of LC3B to autophagosomes, as well as general protein deacetylation. Altogether, these results indicate that coffee triggers 2 phenomena that are also induced by nutrient depletion, namely a reduction of protein acetylation coupled to an increase in autophagy. We speculate that polyphenols contained in coffee promote health by stimulating autophagy. PMID:24769862

  9. In vivo generator for radioimmunotherapy

    DOEpatents

    Mausner, Leonard F.; Srivastava, Suresh G.; Straub, Rita F.

    1988-01-01

    The present invention involves labeling monoclonal antibodies with intermediate half-life radionuclides which decay to much shorter half-life daughters with desirable high energy beta emissions. Since the daughter will be in equilibrium with the parent, it can exert an in-situ tumoricidal effect over a prolonged period in a localized fashion, essentially as an "in-vivo generator". This approach circumvents the inverse relationship between half-life and beta decay energy. Compartmental modeling was used to determine the relative distribution of dose from both parent and daughter nuclei in target and non-target tissues. Actual antibody biodistribution data have been used to fit realistic rate constants for a model containing tumor, blood, and non-tumor compartments. These rate constants were then used in a variety of simulations for two generator systems, Ba-128/Cs-128 (t.sub.1/2 =2.4d/3.6m) and Pd-112/Ag-112 (t.sub.1/2 =0.9d/192m). The results show that higher tumor/background dose ratios may be achievable by virtue of the rapid excretion of a chemically different daughter during the uptake and clearance phases. This modeling also quantitatively demonstrates the favorable impact on activity distribution of a faster monoclonal antibody tumor uptake, especially when the antibody is labeled with a radionuclide with a comparable half-life.

  10. In vivo generator for radioimmunotherapy

    DOEpatents

    Mausner, Leonard F.; Srivastava, Suresh G.; Straub, Rita F.

    1988-11-01

    The present invention involves labeling monoclonal antibodies with intermediate half-life radionuclides which decay to much shorter half-life daughters with desirable high energy beta emissions. Since the daughter will be in equilibrium with the parent, it can exert an in-situ tumoricidal effect over a prolonged period in a localized fashion, essentially as an "in-vivo generator". This approach circumvents the inverse relationship between half-life and beta decay energy. Compartmental modeling was used to determine the relative distribution of dose from both parent and daughter nuclei in target and non-target tissues. Actual antibody biodistribution data have been used to fit realistic rate constants for a model containing tumor, blood, and non-tumor compartments. These rate constants were then used in a variety of simulations for two generator systems, Ba-128/Cs-128 (t.sub.1/2 =2.4d/3.6m) and Pd-112/Ag-112 (t.sub.1/2 =0.9d/192m). The results show that higher tumor/background dose ratios may be achievable by virtue of the rapid excretion of a chemically different daughter during the uptake and clearance phases. This modeling also quantitatively demonstrates the favorable impact on activity distribution of a faster monoclonal antibody tumor uptake, especially when the antibody is labeled with a radionuclide with a comparable half-life.

  11. Issues with dying patients.

    PubMed

    Valent, P

    1978-04-22

    Doctors have the privilege of looking after patients from the moment of birth to the moment of death. Yet, the holistic approach to patients is interfered with by the doctor's role as a warrior against death, where death's everpresent claim on our lives, and its final victory, are ignored. This paper attempts to explore why doctors are in their current position, the mechanisms for ignoring death which are shared by doctors and patients, the nature of the fear of death, and practical implications for the treatment of dying patients. More and more patients die now in medical settings. It is incumbent on doctors to understand the dying process, if much unnecessary suffering is to be prevented. PMID:661717

  12. A Comparison of General Case In Vivo and General Case Simulation Plus In Vivo Training.

    ERIC Educational Resources Information Center

    McDonnell, John J.; Ferguson, Brad

    1988-01-01

    The study examined the relative effectiveness and efficiency of general case in vivo and general case simulation plus in vivo training in teaching six students with moderate and severe disabilities to purchase items in fast-food restaurants. Although both strategies led to reliable performance in nontrained settings, the in vivo instruction…

  13. Experiences of the dying.

    PubMed

    Schoenbeck, Susan L

    2011-01-01

    It is often a mystery to us how we have come to know and believe in certain things. Beliefs are like guests who come up to a door. They come in only if the host opens it and invites them in. Otherwise they are turned away, unable to enter. LPNs/LVNs are invited to reflect on their experiences and expand their knowledge and beliefs. There is growing recognition that bedside talks of the dying, spirit travel and near-death events are real events for the people who experience them. LPNs/ LVNs are encouraged to expand their knowledge and beliefs about dying. PMID:23252027

  14. Assisted Dying in Canada.

    PubMed

    Schuklenk, Udo

    2014-01-01

    This paper makes an affirmative ethical case in favour of the decriminalization of assisted dying in Canada. It then proceeds to defending the affirmative case against various slippery-slope arguments that are typically deployed by opponents of assisted dying. Finally, a recent case of questionable professional conduct by anti-euthanasia campaigners cum academics is flagged as a warning to all of us not to permit the quality of the professional debate to deteriorate unacceptably, despite the personal emotional investments involved on all sides of the debate. PMID:26871530

  15. Absolute calibration in vivo measurement systems

    SciTech Connect

    Kruchten, D.A.; Hickman, D.P.

    1991-02-01

    Lawrence Livermore National Laboratory (LLNL) is currently investigating a new method for obtaining absolute calibration factors for radiation measurement systems used to measure internally deposited radionuclides in vivo. Absolute calibration of in vivo measurement systems will eliminate the need to generate a series of human surrogate structures (i.e., phantoms) for calibrating in vivo measurement systems. The absolute calibration of in vivo measurement systems utilizes magnetic resonance imaging (MRI) to define physiological structure, size, and composition. The MRI image provides a digitized representation of the physiological structure, which allows for any mathematical distribution of radionuclides within the body. Using Monte Carlo transport codes, the emission spectrum from the body is predicted. The in vivo measurement equipment is calibrated using the Monte Carlo code and adjusting for the intrinsic properties of the detection system. The calibration factors are verified using measurements of existing phantoms and previously obtained measurements of human volunteers. 8 refs.

  16. Poetry and the Dying.

    ERIC Educational Resources Information Center

    Kramer, Aaron

    1992-01-01

    Demonstrates roles poetry can play as people confront the death of loved ones and their own dying. Gives examples of Heinrich Heine transforming his agony into art and, from the poetry of two college students, both in advanced stages of neurological disease, which was read aloud in class, teaching all present something about how to approach their…

  17. Tool & Die Technician.

    ERIC Educational Resources Information Center

    Ohio State Univ., Columbus. Center on Education and Training for Employment.

    This document contains 23 units to consider for use in a tech prep competency profile for the occupation of tool and die technician. All the units listed will not necessarily apply to every situation or tech prep consortium, nor will all the competencies within each unit be appropriate. Several units appear within each specific occupation and…

  18. When a Baby Dies.

    ERIC Educational Resources Information Center

    Church, Martha Jo; And Others

    Written especially for grieving mothers whose babies have died, this booklet offers an overview of stages and experiences through which bereaved parents commonly pass. Specifically, the text is intended to give comfort to bereaved parents, offer insight into the grieving process, and provide thoughts on leave-taking ceremonies. The first section…

  19. Tailoring vessel morphology in vivo

    NASA Astrophysics Data System (ADS)

    Gould, Daniel Joseph

    Tissue engineering is a rapidly growing field which seeks to provide alternatives to organ transplantation in order to address the increasing need for transplantable tissues. One huge hurdle in this effort is the provision of thick tissues; this hurdle exists because currently there is no way to provide prevascularized or rapidly vascularizable scaffolds. To design thick, vascularized tissues, scaffolds are needed that can induce vessels which are similar to the microvasculature found in normal tissues. Angiogenic biomaterials are being developed to provide useful scaffolds to address this problem. In this thesis angiogenic and cell signaling and adhesion factors were incorporated into a biomimetic poly(ethylene glycol) (PEG) hydrogel system. The composition of these hydrogels was precisely tuned to induce the formation of differing vessel morphology. To sensitively measure induced microvascular morphology and to compare it to native microvessels in several tissues, this thesis developed an image-based tool for quantification of scale invariant and classical measures of vessel morphology. The tool displayed great utility in the comparison of native vessels and remodeling vessels in normal tissues. To utilize this tool to tune the vessel response in vivo, Flk1::myr-mCherry fluorescently labeled mice were implanted with Platelet Derived Growth Factor-BB (PDGF-BB) and basic Fibroblast Growth Factor (FGF-2) containing PEG-based hydrogels in a modified mouse corneal angiogenesis assay. Resulting vessels were imaged with confocal microscopy, analyzed with the image based tool created in this thesis to compare morphological differences between treatment groups, and used to create a linear relationship between space filling parameters and dose of growth factor release. Morphological parameters of native mouse tissue vessels were then compared to the linear fit to calculate the dose of growth factors needed to induce vessels similar in morphology to native vessels

  20. Navigating "Assisted Dying".

    PubMed

    Schipper, Harvey

    2016-02-01

    Carter is a bellwether decision, an adjudication on a narrow point of law whose implications are vast across society, and whose impact may not be realized for years. Coupled with Quebec's Act Respecting End-of-life Care it has sharply changed the legal landscape with respect to actively ending a person's life. "Medically assisted dying" will be permitted under circumstances, and through processes, which have yet to be operationally defined. This decision carries with it moral assumptions, which mean that it will be difficult to reach a unifying consensus. For some, the decision and Act reflect a modern acknowledgement of individual autonomy. For others, allowing such acts is morally unspeakable. Having opened the Pandora's Box, the question becomes one of navigating a tolerable societal path. I believe it is possible to achieve a workable solution based on the core principle that "medically assisted dying" should be a very rarely employed last option, subject to transparent ongoing review, specifically as to why it was deemed necessary. My analysis is based on 1. The societal conditions in which have fostered demand for "assisted dying", 2. Actions in other jurisdictions, 3. Carter and Quebec Bill 52, 4. Political considerations, 5. Current medical practice. Leading to a series of recommendations regarding. 1. Legislation and regulation, 2. The role of professional regulatory agencies, 3. Medical professions education and practice, 4. Public education, 5. Health care delivery and palliative care. Given the burden of public opinion, and the legal steps already taken, a process for assisted-dying is required. However, those legal and regulatory steps should only be considered a necessary and defensive first step in a two stage process. The larger goal, the second step, is to drive the improvement of care, and thus minimize assisted-dying. PMID:27169205

  1. Herausforderungen durch die deutsche Wiedervereinigung

    NASA Astrophysics Data System (ADS)

    Stäglin, Reiner

    Die Wiedervereinigung stellte auch die Statistik vor große Aufgaben. Die als Organ der staatlichen Planung staatsnah orientierte Statistik der DDR musste auf das zur Neutralität und wissenschaftlichen Unabhängigkeit verpflichtete System der Bundesrepublik umgestellt werden. Ebenso verlangten die Universitäten eine Neuorientierung. Die Deutsche Statistische Gesellschaft hat sich vor allem dreier Aufgaben mit großem Engagement, aber auch mit Bedachtsamkeit angenommen: Aufnahme und Integration der Statistiker aus den neuen Bundesländern in die Gesellschaft, Begleitung der Neuausrichtung des Faches Statistik an deren Hochschulen und Sicherung sowie Nutzung von Datenbeständen der ehemaligen DDR.

  2. Modeling the Mechanical Performance of Die Casting Dies

    SciTech Connect

    R. Allen Miller

    2004-02-27

    The following report covers work performed at Ohio State on modeling the mechanical performance of dies. The focus of the project was development and particularly verification of finite element techniques used to model and predict displacements and stresses in die casting dies. The work entails a major case study performed with and industrial partner on a production die and laboratory experiments performed at Ohio State.

  3. Heated die facilitates tungsten forming

    NASA Technical Reports Server (NTRS)

    Chattin, J. H.; Haystrick, J. E.; Laughlin, J. C.; Leidy, R. A.

    1966-01-01

    Tungsten forming in a press brake employs a bottom die assembly with a heating manifold between two water-cooled die sections. The manifold has hydrogen-oxygen burners spaced along its length for even heat during forming.

  4. Noninvasive photoacoustic microscopy of methemoglobin in vivo

    PubMed Central

    Tang, Min; Zhou, Yong; Zhang, Ruiying; Wang, Lihong V.

    2015-01-01

    Abstract. Due to the various causes of methemoglobinemia and its potential to be confused with other diseases, in vivo measurements of methemoglobin have significant applications in the clinic. Using photoacoustic microscopy (PAM), we quantified the average and the distributed percentage of methemoglobin both in vitro and in vivo. Based on the absorption spectra of methemoglobin, oxyhemoglobin, and deoxyhemoglobin, three wavelengths were chosen to differentiate methemoglobin from the others. The methemoglobin concentrations calculated from the photoacoustic signals agreed well with the preset concentrations. Then we imaged the methemoglobin percentage in microtubes that mimicked blood vessels. Average percentages calculated for five samples with different methemoglobin concentrations also agreed well with the preset values. Finally, we demonstrated the ability of PAM to detect methemoglobin in vivo in a mouse ear. Our results show that PAM can quantitatively image methemoglobin distribution in vivo. PMID:25760655

  5. Genetic targeting of chemical indicators in vivo.

    PubMed

    Yang, Guoying; de Castro Reis, Fernanda; Sundukova, Mayya; Pimpinella, Sofia; Asaro, Antonino; Castaldi, Laura; Batti, Laura; Bilbao, Daniel; Reymond, Luc; Johnsson, Kai; Heppenstall, Paul A

    2015-02-01

    Fluorescent protein reporters have become the mainstay for tracing cellular circuitry in vivo but are limited in their versatility. Here we generated Cre-dependent reporter mice expressing the Snap-tag to target synthetic indicators to cells. Snap-tag labeling worked efficiently and selectively in vivo, allowing for both the manipulation of behavior and monitoring of cellular fluorescence from the same reporter. PMID:25486061

  6. Psychotherapy with Older Dying Persons.

    ERIC Educational Resources Information Center

    Dye, Carol J.

    Psychotherapy with older dying patients can lead to problems of countertransference for the clinician. Working with dying patients requires flexibility to adapt basic therapeutics to the institutional setting. Goals of psychotherapy must be reconceptualized for dying clients. The problems of countertransference arise because clinicians themselves…

  7. In vivo studies of opiate receptors

    SciTech Connect

    Frost, J.J.; Dannals, R.F.; Duelfer, T.; Burns, H.D.; Ravert, H.T.; Langstroem, B.; Balasubramanian, V.; Wagner, H.N. Jr.

    1984-01-01

    To study opiate receptors noninvasively in vivo using positron emission tomography, techniques for preferentially labeling opiate receptors in vivo can be used. The rate at which receptor-bound ligand clears from the brain in vivo can be predicted by measuring the equilibrium dissociation constant (KD) at 37 degrees C in the presence of 100 mM sodium chloride and 100 microM guanyl-5'-imidodiphosphate, the drug distribution coefficient, and the molecular weight. A suitable ligand for labeling opiate receptors in vivo is diprenorphine, which binds to mu, delta, and kappa receptors with approximately equal affinity in vitro. However, in vivo diprenorphine may bind predominantly to one opiate receptor subtype, possibly the mu receptor. To predict the affinity for binding to the opiate receptor, a Hansch correlation was determined between the 50% inhibitory concentration for a series of halogen-substituted fentanyl analogs and electronic, lipophilic, and steric parameters. Radiochemical methods for the synthesis of carbon-11-labeled diprenorphine and lofentanil are presented.

  8. Nitriding of Aluminum Extrusion Die: Effect of Die Geometry

    NASA Astrophysics Data System (ADS)

    Akhtar, S. S.; Arif, A. F. M.; Yilbas, B. S.

    2010-04-01

    Nitriding of complex-shaped extrusion dies may result in non-uniform nitride layers and hence a required hardness may not be achieved in some regions of the bearing area. The present study is carried out to assess the effect of extrusion die profile on the characteristics and growth behavior of nitride layers so that the critical die design feature can be identified to enhance the uniformity of the nitride layer. For this purpose, AISI H13 steel samples have been manufactured with profiles similar to those of hot extrusion dies. The samples were then gas nitrided under controlled nitriding potential. The uniformity and depth of nitride layers have been investigated in terms of compound layer and total nitride case depth for selected die features. The results of this study indicated the need to include the effect of profile on the nitride layer for the optimal die design with improved service life.

  9. Accurate defect die placement and nuisance defect reduction for reticle die-to-die inspections

    NASA Astrophysics Data System (ADS)

    Wen, Vincent; Huang, L. R.; Lin, C. J.; Tseng, Y. N.; Huang, W. H.; Tuo, Laurent C.; Wylie, Mark; Chen, Ellison; Wang, Elvik; Glasser, Joshua; Kelkar, Amrish; Wu, David

    2015-10-01

    Die-to-die reticle inspections are among the simplest and most sensitive reticle inspections because of the use of an identical-design neighboring-die for the reference image. However, this inspection mode can have two key disadvantages: (1) The location of the defect is indeterminate because it is unclear to the inspector whether the test or reference image is defective; and (2) nuisance and false defects from mask manufacturing noise and tool optical variation can limit the usable sensitivity. The use of a new sequencing approach for a die-to-die inspection can resolve these issues without any additional scan time, without sacrifice in sensitivity requirement, and with a manageable increase in computation load. In this paper we explore another approach for die-to-die inspections using a new method of defect processing and sequencing. Utilizing die-to-die double arbitration during defect detection has been proven through extensive testing to generate accurate placement of the defect in the correct die to ensure efficient defect disposition at the AIMS step. The use of this method maintained the required inspection sensitivity for mask quality as verified with programmed-defectmask qualification and then further validated with production masks comparing the current inspection approach to the new method. Furthermore, this approach can significantly reduce the total number of defects that need to be reviewed by essentially eliminating the nuisance and false defects that can result from a die-to-die inspection. This "double-win" will significantly reduce the effort in classifying a die-to-die inspection result and will lead to improved cycle times.

  10. Dissolution DNP for in vivo preclinical studies

    NASA Astrophysics Data System (ADS)

    Comment, Arnaud

    2016-03-01

    The tremendous polarization enhancement afforded by dissolution dynamic nuclear polarization (DNP) can be taken advantage of to perform preclinical in vivo molecular and metabolic imaging. Following the injection of molecules that are hyperpolarized via dissolution DNP, real-time measurements of their biodistribution and metabolic conversion can be recorded. This technology therefore provides a unique and invaluable tool for probing cellular metabolism in vivo in animal models in a noninvasive manner. It gives the opportunity to follow and evaluate disease progression and treatment response without requiring ex vivo destructive tissue assays. Although its considerable potential has now been widely recognized, hyperpolarized magnetic resonance by dissolution DNP remains a challenging method to implement for routine in vivo preclinical measurements. The aim of this article is to provide an overview of the current state-of-the-art technology for preclinical applications and the challenges that need to be addressed to promote it and allow its wider dissemination in the near future.

  11. In Vivo Production of Entomopathogenic Nematodes.

    PubMed

    Shapiro-Ilan, David I; Morales-Ramos, Juan A; Rojas, M Guadalupe

    2016-01-01

    In nature, entomopathogenic nematodes in the genera Heterorhabditis and Steinernema are obligate parasites of insects. The nematodes are used widely as biopesticides for suppression of insect pests. More than a dozen entomopathogenic nematode species have been commercialized for use in biological control. Most nematodes intended for commercial application are produced in artificial media via solid or liquid fermentation. However, for laboratory research and small greenhouse or field trials, in vivo production of entomopathogenic nematodes is the common method of propagation. Additionally, small companies continue to produce nematodes using in vivo methods for application in niche markets. Advances in mechanization and alternative production routes (e.g., production geared toward application of nematodes in infected host cadavers) can improve efficiency and economy of scale. The objective of this chapter is to describe basic and advanced procedures for in vivo production of entomopathogenic nematodes. PMID:27565497

  12. Grueneisen relaxation photoacoustic microscopy in vivo

    NASA Astrophysics Data System (ADS)

    Ma, Jun; Shi, Junhui; Hai, Pengfei; Zhou, Yong; Wang, Lihong V.

    2016-06-01

    Grueneisen relaxation photoacoustic microscopy (GR-PAM) can achieve optically defined axial resolution, but it has been limited to ex vivo demonstrations so far. Here, we present the first in vivo image of a mouse brain acquired with GR-PAM. To induce the GR effect, an intensity-modulated continuous-wave laser was employed to heat absorbing objects. In phantom experiments, an axial resolution of 12.5 μm was achieved, which is sixfold better than the value achieved by conventional optical-resolution PAM. This axial-resolution improvement was further demonstrated by imaging a mouse brain in vivo, where significantly narrower axial profiles of blood vessels were observed. The in vivo demonstration of GR-PAM shows the potential of this modality for label-free and high-resolution anatomical and functional imaging of biological tissues.

  13. Dissolution DNP for in vivo preclinical studies.

    PubMed

    Comment, Arnaud

    2016-03-01

    The tremendous polarization enhancement afforded by dissolution dynamic nuclear polarization (DNP) can be taken advantage of to perform preclinical in vivo molecular and metabolic imaging. Following the injection of molecules that are hyperpolarized via dissolution DNP, real-time measurements of their biodistribution and metabolic conversion can be recorded. This technology therefore provides a unique and invaluable tool for probing cellular metabolism in vivo in animal models in a noninvasive manner. It gives the opportunity to follow and evaluate disease progression and treatment response without requiring ex vivo destructive tissue assays. Although its considerable potential has now been widely recognized, hyperpolarized magnetic resonance by dissolution DNP remains a challenging method to implement for routine in vivo preclinical measurements. The aim of this article is to provide an overview of the current state-of-the-art technology for preclinical applications and the challenges that need to be addressed to promote it and allow its wider dissemination in the near future. PMID:26920829

  14. Outer Hair Cell Electromotility in vivo

    NASA Astrophysics Data System (ADS)

    Ramamoorthy, Sripriya; Nuttall, Alfred L.

    2011-11-01

    The effectiveness of outer hair cell (OHC) electro-motility in vivo has been challenged by the expected low-pass filtering of the transmembrane potential due to the cell's own capacitance. The OHC electromotility is characterized here by an electromechanical ratio defined as the ratio of the OHC contraction to the transmembrane potential. This ratio has been measured in isolated cells to be approximately 26 nm/mV. We estimate the OHC electromechanical ratio in vivo from the recently measured displacements of the reticular lamina and the basilar membrane near the 19 kHz characteristic frequency in the basal region of guinea pig cochlea. Our analysis strongly suggests OHC electromotility process is effective for cochlear amplification in vivo at least around the characteristic frequency of the basal location in spite of the low-pass filtering.

  15. In vivo fluorescence lifetime optical projection tomography

    PubMed Central

    McGinty, James; Taylor, Harriet B.; Chen, Lingling; Bugeon, Laurence; Lamb, Jonathan R.; Dallman, Margaret J.; French, Paul M. W.

    2011-01-01

    We demonstrate the application of fluorescence lifetime optical projection tomography (FLIM-OPT) to in vivo imaging of lysC:GFP transgenic zebrafish embryos (Danio rerio). This method has been applied to unambiguously distinguish between the fluorescent protein (GFP) signal in myeloid cells from background autofluorescence based on the fluorescence lifetime. The combination of FLIM, an inherently ratiometric method, in conjunction with OPT results in a quantitative 3-D tomographic technique that could be used as a robust method for in vivo biological and pharmaceutical research, for example as a readout of Förster resonance energy transfer based interactions. PMID:21559145

  16. Graphite/Thermoplastic-Pultrusion Die

    NASA Technical Reports Server (NTRS)

    Wilson, Maywood L.; Frye, Mark W.; Johnson, Gary S.; Stanfield, Clarence E.

    1990-01-01

    Attachment to extruder produces thermoplastic-impregnated graphite tape. Consists of profile die, fiber/resin collimator, and crosshead die body. Die designed to be attached to commercially available extrusion machine capable of extruding high-performance thermoplastics. Simple attachment to commercial extruder enables developers of composites to begin experimenting with large numbers of proprietary resins, fibers, and hybrid composite structures. With device, almost any possible fiber/resin combination fabricated.

  17. Foreigners dying in Istanbul.

    PubMed

    Uzun, Ibrahim; Celbis, Osman; Baydar, Cetin Lutfi; Alkan, Nevzat; Arslan, Murat Nihat

    2009-09-01

    The study included 411 deaths selected from 14,647 medicolegal deaths autopsied in the Morgue Department of Forensic Medicine Institute Directorate, affiliated with the Ministry of Justice, between 1998 and 2002. Data were collected from court documents, coroner's investigation reports, and autopsy reports. The parameters of age, gender, nationality and origin, cause and place of death in foreigners dying in Istanbul were evaluated in the study. Out of 14,647 medicolegal deaths, 3.5% were foreigners from 34 different nationalities. The nationality with the highest rate of foreigner deaths (34%) was Romanian. Out of 411 deaths, 74.3% were male and 25.7% were female. Of all cases, 64.4% were tourists visiting Istanbul and 35.6% had a job in Istanbul. Of 146 foreigners employed in Istanbul, 94.5% did not have a work permit, while only 5.5% had a work permit. PMID:19674242

  18. Toward Localized In Vivo Biomarker Concentration Measurements

    PubMed Central

    Zhang, Xiaojuan; Reeves, Daniel; Shi, Yipeng; Gimi, Barjor; Nemani, Krishnamurthy V.; Perreard, Irina M.; Toraya-Brown, Seiko; Fiering, Steven; Weaver, John B.

    2015-01-01

    We know a great deal about the biochemistry of cells because they can be isolated and studied. The biochemistry of the much more complex in vivo environment is more difficult to study because the only ways to quantitate concentrations is to sacrifice the animal or biopsy the tissue. Either method disrupts the environment profoundly and neither method allows longitudinal studies on the same individual. Methods of measuring chemical concentrations in vivo are very valuable alternatives to sacrificing groups of animals. We are developing microscopic magnetic nanoparticle (mNP) probes to measure the concentration of a selected molecule in vivo. The mNPs are targeted to bind the selected molecule and the resulting reduction in rotational freedom can be quantified remotely using magnetic spectroscopy. The mNPs must be contained in micrometer sized porous shells to keep them from migrating and to protect them from clearance by the immune system. There are two key issues in the development of the probes. First, we demonstrate the ability to measure concentrations in the porous walled alginate probes both in phosphate buffered saline and in blood, which is an excellent surrogate for the complex and challenging in vivo environment. Second, sensitivity is critical because it allows microscopic probes to measure very small concentrations very far away. We report sensitivity measurements on recently introduced technology that has allowed us to improve the sensitivity by two orders of magnitude, a factor of 200 so far. PMID:26203196

  19. HIV control in vivo: Dynamical analysis

    NASA Astrophysics Data System (ADS)

    Gumel, A. B.; Moghadas, S. M.

    2004-10-01

    A deterministic model for the immunological and therapeutic control of human immunodeficiency virus (HIV) in vivo is studied qualitatively. In addition to analyzing the local stability of the equilibria, the global stability of the infection-free equilibrium is established. The optimal efficacy level of anti-retroviral therapy needed to eradicate HIV from the body of an HIV-infected individual is obtained.

  20. In vivo dosimetry in external beam radiotherapy

    SciTech Connect

    Mijnheer, Ben; Beddar, Sam; Izewska, Joanna; Reft, Chester

    2013-07-15

    In vivo dosimetry (IVD) is in use in external beam radiotherapy (EBRT) to detect major errors, to assess clinically relevant differences between planned and delivered dose, to record dose received by individual patients, and to fulfill legal requirements. After discussing briefly the main characteristics of the most commonly applied IVD systems, the clinical experience of IVD during EBRT will be summarized. Advancement of the traditional aspects of in vivo dosimetry as well as the development of currently available and newly emerging noninterventional technologies are required for large-scale implementation of IVD in EBRT. These new technologies include the development of electronic portal imaging devices for 2D and 3D patient dosimetry during advanced treatment techniques, such as IMRT and VMAT, and the use of IVD in proton and ion radiotherapy by measuring the decay of radiation-induced radionuclides. In the final analysis, we will show in this Vision 20/20 paper that in addition to regulatory compliance and reimbursement issues, the rationale for in vivo measurements is to provide an accurate and independent verification of the overall treatment procedure. It will enable the identification of potential errors in dose calculation, data transfer, dose delivery, patient setup, and changes in patient anatomy. It is the authors' opinion that all treatments with curative intent should be verified through in vivo dose measurements in combination with pretreatment checks.

  1. In-vivo optical investigation of psoriasis

    NASA Astrophysics Data System (ADS)

    Kapsokalyvas, Dimitrios; Cicchi, Riccardo; Bruscino, Nicola; Alfieri, Domenico; Massi, Daniela; Lotti, Torello; Pavone, Francesco S.

    2011-03-01

    Psoriasis is an autoimmune disease of the skin characterized by hyperkeratosis, hyperproliferation of the epidermis, inflammatory cell accumulation and increased dilatation of dermal papillary blood vessels. Cases of psoriasis were investigated in vivo with optical means in order to evaluate the potential of in vivo optical biopsy. A Polarization Multispectral Dermoscope was employed for the macroscopic observation. Features such as the 'dotted' blood vessels pattern was observed with high contrast. The average size of dot vessels in Psoriasis was measured to be 974 μm2 which is much higher compared to healthy skin. High resolution image sections of the epidermis and the dermis were produced with a custom made Multiphoton Microscope. Imaging extended from the surface of the lesion down to the papillary dermis, at a depth of 200 μm. In the epidermis, a characteristic morphology of the stratum corneum found only in Psoriasis was revealed. Additionally, the cytoplasmic area of the cells in the stratum spinosum layer was found to be smaller than normal. In the dermis the morphological features were more pronounced, where the elongated dermal papillae dominated the papillary layer. Their length exceeds 100μm, which is a far greater value compared to that of healthy skin. These in vivo observations are consistent with the ex vivo histopathological observations, supporting both the applicability and potentiality of multispectral dermoscopy and multiphoton microscopy in the field of in vivo optical investigation and biopsy of skin.

  2. Two Piece Compaction Die Design

    SciTech Connect

    Coffey, Ethan N

    2010-03-01

    Compaction dies used to create europium oxide and tantalum control plates were modeled using ANSYS 11.0. Two-piece designs were considered in order to make the dies easier to assemble than the five-piece dies that were previously used. The two areas of concern were the stresses at the interior corner of the die cavity and the distortion of the cavity wall due to the interference fit between the two pieces and the pressure exerted on the die during the compaction process. A successful die design would have stresses less than the yield stress of the material and a maximum wall distortion on the order of 0.0001 in. Design factors that were investigated include the inner corner radius, the value of the interference fit, the compaction force, the size of the cavity, and the outer radius and geometry of the outer ring. The results show that for the europium oxide die, a 0.01 in. diameter wire can be used to create the cavity, leading to a 0.0055 in. radius corner, if the radial interference fit is 0.003 in. For the tantalum die, the same wire can be used with a radial interference fit of 0.001 in. Also, for the europium oxide die with a 0.003 in. interference fit, it is possible to use a wire with a diameter of 0.006 in. for the wire burning process. Adding a 10% safety factor to the compaction force tends to lead to conservative estimates of the stresses but not for the wall distortion. However, when the 10% safety factor is removed, the wall distortion is not affected enough to discard the design. Finally, regarding the europium oxide die, when the cavity walls are increased by 0.002 in. per side or the outer ring is made to the same geometry as the tantalum die, all the stresses and wall distortions are within the desired range. Thus, the recommendation is to use a 0.006 in. diameter wire and a 0.003 in. interference fit for the europium oxide die and a 0.01 in. diameter wire and a 0.001 in. interference fit for the tantalum die. The dies can also be made to have the

  3. Adapt or die?

    PubMed

    Visser, S S; Nel, A H

    1996-12-01

    The worldwide economic recession and the concomitant limited stock of finances have had an influence on the available money of every household and have also inhibited the improvement of socio-economic conditions and medicine. The Reconstruction and Development Programme (RDP) has the objective of improving the living conditions of the people with regard to housing, education, training and health care. The latter seems to be a major problem which has to be addressed with the emphasis on the preventive and promotional aspects of health care. A comprehensive health care system did not come into being property in the past because of the maldistribution of health care services, personnel and differences in culture and health care beliefs and values. The question that now arises, is how to render a quality health care service within the constraints of inadequate financing and resources. A comprehensive literature study has been done with reference to quality health care and financing followed by a survey of existing health services and finances. Recommendations are made about minimum requirements to be accepted if one were to adapt rather than die in terms of the provision of healthcare: the decentralization and rationalization of the administration of health care, the stress on and realization of effective and efficient primary health care, the acceptance of participative management in health providing organizations, the provision of financial management training for health care managers and the application of management accounting principles for the improvement of the efficiency and effectiveness of management. PMID:9283343

  4. Dying and multiplying life.

    PubMed

    Rodríguez-Arias, David

    2014-09-01

    It was only after James P. Lovette's death, in 2006, that I discovered that the twenty-four-year-old colleague and friend with whom I had spent so many afternoons debating issues in organ transplantation had been the first successful child heart transplantee in the world and one of the longest-living survivors of a second transplant. During the years we met, he never even hinted at the fact that three different hearts had beaten in his chest. The revelation that his life had been an almost uninterrupted chain of medical challenges suddenly made me appreciate his quirkiness in a whole new light. Organ transplantation crudely exemplifies a traditional moral dilemma between means and ends: in order to save a life, someone else has to die. Bioethicists involved in this field have the role of identifying the ethical issues surrounding organ donation and helping others to argue in an intelligible and convincing way. In my view, bioethicists have the obligation to foster a discussion as open and transparent as possible on these matters. Still, I sometimes fear that I may be helping to cause unnecessary harms to potential recipients who are desperately waiting for a vital organ. Scholars would be chillingly cold if their quest for truth systematically came at the cost of lives lost. Every life can be meaningful and provide meaning to many others. This is true even with organ recipients, who often have short lives full of considerable suffering. PMID:25231665

  5. Chitosan mouthwash: toxicity and in vivo validation.

    PubMed

    Costa, E M; Silva, S; Costa, M R; Pereira, M; Campos, D A; Odila, J; Madureira, A R; Cardelle-Cobas, A; Tavaria, F K; Rodrigues, A S; Pintado, M M

    2014-10-13

    A previous study showed that a chitosan mouthwash would be a valid alternative to current mouthwashes as it demonstrated, in vitro, significantly higher antibiofilm activity than two commercial mouthwashes. As such, the aim of this work was to verify the safety of the developed product and to validate, in vivo, the biological activity ascertained in vitro. Chitosan mouthwash safety was evaluated through Ames, MTT and V79 chromosomal aberration assay while antimicrobial activity was evaluated through in vivo assays. The results showed that the chitosan mouthwash was safe, presenting lower cytotoxicity than a commercial mouthwash, and that it effectively reduced viable counts of Streptococcus spp. and Enterococcus spp. by ca. 5.5 log of CFU. Furthermore, in direct comparison with a commercial mouthwash the chitosan mouthwash possessed significantly higher antimicrobial activity. The conjunction of these results proves that the chitosan mouthwash is a safe, effective, natural alternative to the existent chemical mouthwashes. PMID:25037365

  6. Preconditioning Stem Cells for In Vivo Delivery

    PubMed Central

    Sart, Sébastien; Ma, Teng

    2014-01-01

    Abstract Stem cells have emerged as promising tools for the treatment of incurable neural and heart diseases and tissue damage. However, the survival of transplanted stem cells is reported to be low, reducing their therapeutic effects. The major causes of poor survival of stem cells in vivo are linked to anoikis, potential immune rejection, and oxidative damage mediating apoptosis. This review investigates novel methods and potential molecular mechanisms for stem cell preconditioning in vitro to increase their retention after transplantation in damaged tissues. Microenvironmental preconditioning (e.g., hypoxia, heat shock, and exposure to oxidative stress), aggregate formation, and hydrogel encapsulation have been revealed as promising strategies to reduce cell apoptosis in vivo while maintaining biological functions of the cells. Moreover, this review seeks to identify methods of optimizing cell dose preparation to enhance stem cell survival and therapeutic function after transplantation. PMID:25126478

  7. Evaluation of vaginal antifungal formulations in vivo.

    PubMed Central

    McRipley, R. J.; Erhard, P. J.; Schwind, R. A.; Whitney, R. R.

    1979-01-01

    Relatively simple and rapid procedures have been developed for evaluating the local efficacy of vaginal antifungal agents in vivo in a vaginal candidiasis model in ovariectomized rats. The results of this investigation indicate that the model and methods described are quite suitable for screening potential antifungal substances and for assessing the chemotherapeutic effectiveness of new antifungal agents and formulations before carrying out clinical studies. PMID:392480

  8. Laser spectrofluorometry of agronomic plants in vivo

    NASA Astrophysics Data System (ADS)

    Posudin, Yuri I.

    1997-05-01

    Laser spectrofluorometry of agronomic plants in vivo at the single leaf level permits to investigate the effects of the growth phase, part, nodal position and age of the leaf, agrochemical treatment, external physical factors and plant diseases on the chlorophyll fluorescence. Such a spectroscopic approach can give the possibility to monitoring the development and health status of agronomic plants during ontogenesis and under different stresses. The relevant spectral wavelengths and criteria which correlate with status of the plant were determined.

  9. Photosensitizer quantitation in vivo by flourescence microsampling

    NASA Astrophysics Data System (ADS)

    Pogue, Brian W.; Burke, Gregory C.; Lee, Claudia C.; Hoopes, P. Jack

    2000-06-01

    Photodynamic therapy can provide a reliable method of tumor destruction when the appropriate dosimetry is applied. Current dosimetry practice involves quantification of the drug and light doses applied to the tumor, but it would be desirable to monitor in vivo light and drug levels to provide the most accurate determination of dosimetry. In vivo measurements can be used to minimize variations in treatment response due to inter-animal variability, by providing animal-specific or patient-specific treatment planning. This study reports on the development of a micro-sampling method to measure fluorescence from tissue, which is not significantly affected by the tissue optical properties. The system measures fluorescence from the surface of a tissue, using a fiber bundle composed of individual 100 micron fibers which ar all spaced apart by 700 microns from one another at the tissue contact end. This design provides sampling of the fluorescence at multiple sites to increase the signal intensity, while maintaining a micro- sampling of the tissue volume just below the surface. The calibration studies here indicate that the 1/e sampling depth is near 60 microns when measured in optical phantoms, which are similar to typical tissue properties. The probe fluorescence signal is independent of blood concentration up to a maximum of 10% blood by volume, which is similar to most tumor tissue. Animal tests indicate that the sensitivity to drug concentration is essentially the same in when measured in murine liver and muscle tissues, both in vivo and ex vivo. These preliminary calibration results suggest that the probe can be used to measure photosensitizer uptake in vivo non- invasively and rapidly via conversion of fluorescence intensity to photosensitizer concentration.

  10. In vivo imaging of neocortical epilepsy

    NASA Astrophysics Data System (ADS)

    Schwartz, Theodore

    2003-03-01

    Epilepsy is a disease affecting 1-2Electrical recordings from chronic animal models and human neocortical epileptic foci indicate that the population of neurons underlying each interictal epileptiform discharge varies over time. The spatial relationship between interictal events and the ictal onset zone, thought to be the critical area of epileptogenesis, is not well understood and critical to the surgical treatment of epilepsy. Electrophysiological recording methods, although currently the "gold standard", are inadequate to address these questions based on restrictions due to volume conduction or sampling limitations, many of which can be overcome with optical recording techniques. In vivo optical recording of intrinsic signals can be used to generate high-resolution, real-time maps of the population of neurons participating in an epileptiform event. The goal of our laboratory is to examine the shifting spatio-temporal dynamics of the epileptogenic aggregate in both acute and chronic experimental models of in vivo rodent epilepsy. In particular we are interested in the precise relationship between the optical signal and the interictal and ictal epileptiform events using well-established acute and chronic in vivo rodent models. Optical epilepsy maps recorded at various wavelengths are correlated with maps derived from electrophysiological recordings from multiple surface electrodes.

  11. Laser Nanosurgery of Cerebellar Axons In Vivo

    PubMed Central

    Allegra Mascaro, Anna L.; Sacconi, Leonardo; Pavone, Francesco Saverio

    2014-01-01

    Only a few neuronal populations in the central nervous system (CNS) of adult mammals show local regrowth upon dissection of their axon. In order to understand the mechanism that promotes neuronal regeneration, an in-depth analysis of the neuronal types that can remodel after injury is needed. Several studies showed that damaged climbing fibers are capable of regrowing also in adult animals1,2. The investigation of the time-lapse dynamics of degeneration and regeneration of these axons within their complex environment can be performed by time-lapse two-photon fluorescence (TPF) imaging in vivo3,4. This technique is here combined with laser surgery, which proved to be a highly selective tool to disrupt fluorescent structures in the intact mouse cortex5-9. This protocol describes how to perform TPF time-lapse imaging and laser nanosurgery of single axonal branches in the cerebellum in vivo. Olivocerebellar neurons are labeled by anterograde tracing with a dextran-conjugated dye and then monitored by TPF imaging through a cranial window. The terminal portion of their axons are then dissected by irradiation with a Ti:Sapphire laser at high power. The degeneration and potential regrowth of the damaged neuron are monitored by TPF in vivo imaging during the days following the injury. PMID:25146130

  12. Cells in Dengue Virus Infection In Vivo

    PubMed Central

    Noisakran, Sansanee; Onlamoon, Nattawat; Songprakhon, Pucharee; Hsiao, Hui-Mien; Chokephaibulkit, Kulkanya; Perng, Guey Chuen

    2010-01-01

    Dengue has been recognized as one of the most important vector-borne emerging infectious diseases globally. Though dengue normally causes a self-limiting infection, some patients may develop a life-threatening illness, dengue hemorrhagic fever (DHF)/dengue shock syndrome (DSS). The reason why DHF/DSS occurs in certain individuals is unclear. Studies in the endemic regions suggest that the preexisting antibodies are a risk factor for DHF/DSS. Viremia and thrombocytopenia are the key clinical features of dengue virus infection in patients. The amounts of virus circulating in patients are highly correlated with severe dengue disease, DHF/DSS. Also, the disturbance, mainly a transient depression, of hematological cells is a critical clinical finding in acute dengue patients. However, the cells responsible for the dengue viremia are unresolved in spite of the intensive efforts been made. Dengue virus appears to replicate and proliferate in many adapted cell lines, but these in vitro properties are extremely difficult to be reproduced in primary cells or in vivo. This paper summarizes reports on the permissive cells in vitro and in vivo and suggests a hematological cell lineage for dengue virus infection in vivo, with the hope that a new focus will shed light on further understanding of the complexities of dengue disease. PMID:22331984

  13. Shigella impairs T lymphocyte dynamics in vivo

    PubMed Central

    Salgado-Pabón, Wilmara; Celli, Susanna; Arena, Ellen T.; Nothelfer, Katharina; Roux, Pascal; Sellge, Gernot; Frigimelica, Elisabetta; Bousso, Philippe; Sansonetti, Philippe J.; Phalipon, Armelle

    2013-01-01

    The Gram-negative enteroinvasive bacterium Shigella flexneri is responsible for the endemic form of bacillary dysentery, an acute rectocolitis in humans. S. flexneri uses a type III secretion system to inject effector proteins into host cells, thus diverting cellular functions to its own benefit. Protective immunity to reinfection requires several rounds of infection to be elicited and is short-lasting, suggesting that S. flexneri interferes with the priming of specific immunity. Considering the key role played by T-lymphocyte trafficking in priming of adaptive immunity, we investigated the impact of S. flexneri on T-cell dynamics in vivo. By using two-photon microscopy to visualize bacterium–T-cell cross-talks in the lymph nodes, where the adaptive immunity is initiated, we provide evidence that S. flexneri, via its type III secretion system, impairs the migration pattern of CD4+ T cells independently of cognate recognition of bacterial antigens. We show that bacterial invasion of CD4+ T lymphocytes occurs in vivo, and results in cell migration arrest. In the absence of invasion, CD4+ T-cell migration parameters are also dramatically altered. Signals resulting from S. flexneri interactions with subcapsular sinus macrophages and dendritic cells, and recruitment of polymorphonuclear cells are likely to contribute to this phenomenon. These findings indicate that S. flexneri targets T lymphocytes in vivo and highlight the role of type III effector secretion in modulating host adaptive immune responses. PMID:23417297

  14. What Happens When Someone Dies?

    MedlinePlus

    ... and sleepiness Mental confusion Constipation or incontinence Nausea Refusal to eat or drink Each of these symptoms, ... having a "non-hospital DNR" (see Understanding Health Care Decisions ) if the person is dying at home. ...

  15. REFRACTORY DIE FOR EXTRUDING URANIUM

    DOEpatents

    Creutz, E.C.

    1959-08-11

    A die is presented for the extrusion of metals, said die being formed of a refractory complex oxide having the composition M/sub n/O/sub m/R/sub x/O/sub y/ where M is magnesium, zinc, manganese, or iron, R is aluminum, chromic chromium, ferric iron, or manganic manganese, and m, n, x, and y are whole numbers. Specific examples are spinel, magnesium aluminate, magnetite, magnesioferrite, chromite, and franklinite.

  16. Improving care of dying children.

    PubMed Central

    Martinson, I M

    1995-01-01

    Every year about 5,000 children aged 0 to 14 years need hospice care in the United States. Children seem to know that they are dying, although this is difficult for parents to accept. Clear, empathic understanding is needed. Communication with clarity and understanding is imperative with the changes in goals from cure to palliation to comfort. The ideal place for most dying children is at home, where symptoms can be managed as effectively as in a hospital. PMID:7571589

  17. Unexpected requirement for ELMO1 in clearance of apoptotic germ cells in vivo.

    PubMed

    Elliott, Michael R; Zheng, Shuqiu; Park, Daeho; Woodson, Robin I; Reardon, Michael A; Juncadella, Ignacio J; Kinchen, Jason M; Zhang, Jun; Lysiak, Jeffrey J; Ravichandran, Kodi S

    2010-09-16

    Apoptosis and the subsequent clearance of dying cells occurs throughout development and adult life in many tissues. Failure to promptly clear apoptotic cells has been linked to many diseases. ELMO1 is an evolutionarily conserved cytoplasmic engulfment protein that functions downstream of the phosphatidylserine receptor BAI1, and, along with DOCK1 and the GTPase RAC1, promotes internalization of the dying cells. Here we report the generation of ELMO1-deficient mice, which we found to be unexpectedly viable and grossly normal. However, they had a striking testicular pathology, with disrupted seminiferous epithelium, multinucleated giant cells, uncleared apoptotic germ cells and decreased sperm output. Subsequent in vitro and in vivo analyses revealed a crucial role for ELMO1 in the phagocytic clearance of apoptotic germ cells by Sertoli cells lining the seminiferous epithelium. The engulfment receptor BAI1 and RAC1 (upstream and downstream of ELMO1, respectively) were also important for Sertoli-cell-mediated engulfment. Collectively, these findings uncover a selective requirement for ELMO1 in Sertoli-cell-mediated removal of apoptotic germ cells and make a compelling case for a relationship between engulfment and tissue homeostasis in vivo. PMID:20844538

  18. Could magnetic resonance provide in vivo histology?

    PubMed Central

    Dominietto, Marco; Rudin, Markus

    2014-01-01

    The diagnosis of a suspected tumor lesion faces two basic problems: detection and identification of the specific type of tumor. Radiological techniques are commonly used for the detection and localization of solid tumors. Prerequisite is a high intrinsic or enhanced contrast between normal and neoplastic tissue. Identification of the tumor type is still based on histological analysis. The result depends critically on the sampling sites, which given the inherent heterogeneity of tumors, constitutes a major limitation. Non-invasive in vivo imaging might overcome this limitation providing comprehensive three-dimensional morphological, physiological, and metabolic information as well as the possibility for longitudinal studies. In this context, magnetic resonance based techniques are quite attractive since offer at the same time high spatial resolution, unique soft tissue contrast, good temporal resolution to study dynamic processes and high chemical specificity. The goal of this paper is to review the role of magnetic resonance techniques in characterizing tumor tissue in vivo both at morphological and physiological levels. The first part of this review covers methods, which provide information on specific aspects of tumor phenotypes, considered as indicators of malignancy. These comprise measurements of the inflammatory status, neo-vascular physiology, acidosis, tumor oxygenation, and metabolism together with tissue morphology. Even if the spatial resolution is not sufficient to characterize the tumor phenotype at a cellular level, this multiparametric information might potentially be used for classification of tumors. The second part discusses mathematical tools, which allow characterizing tissue based on the acquired three-dimensional data set. In particular, methods addressing tumor heterogeneity will be highlighted. Finally, we address the potential and limitation of using MRI as a tool to provide in vivo tissue characterization. PMID:24454320

  19. In Vivo Nanodetoxication for Acute Uranium Exposure.

    PubMed

    Guzmán, Luis; Durán-Lara, Esteban F; Donoso, Wendy; Nachtigall, Fabiane M; Santos, Leonardo S

    2015-01-01

    Accidental exposure to uranium is a matter of concern, as U(VI) is nephrotoxic in both human and animal models, and its toxicity is associated to chemical toxicity instead of radioactivity. We synthesized different PAMAM G4 and G5 derivatives in order to prove their interaction with uranium and their effect on the viability of red blood cells in vitro. Furthermore, we prove the effectiveness of the selected dendrimers in an animal model of acute uranium intoxication. The dendrimer PAMAM G4-Lys-Fmoc-Cbz demonstrated the ability to chelate the uranyl ion in vivo, improving the biochemical and histopathologic features caused by acute intoxication with uranium. PMID:26083036

  20. Methods of in vivo radiation measurement

    DOEpatents

    Huffman, Dennis D.; Hughes, Robert C.; Kelsey, Charles A.; Lane, Richard; Ricco, Antonio J.; Snelling, Jay B.; Zipperian, Thomas E.

    1990-01-01

    Methods of and apparatus for in vivo radiation measurements relay on a MOSFET dosimeter of high radiation sensitivity with operates in both the passive mode to provide an integrated dose detector and active mode to provide an irradiation rate detector. A compensating circuit with a matched unirradiated MOSFET is provided to operate at a current designed to eliminate temperature dependence of the device. Preferably, the MOSFET is rigidly mounted in the end of a miniature catheter and the catheter is implanted in the patient proximate the radiation source.

  1. In vivo friction properties of human skin.

    PubMed

    Zhang, M; Mak, A F

    1999-08-01

    In vivo frictional properties of human skin and five materials, namely aluminium, nylon, silicone, cotton sock, Pelite, were investigated. Normal and untreated skin over six anatomic regions of ten normal subjects were measured under a controlled environment. The average coefficient of friction for all measurements is 0.46+/-0.15 (p<0.05). Among all measured sites, the palm of the hand has the highest coefficient of friction (0.62+/-0.22). For all the materials tested, silicone has the highest coefficient of friction (0.61+/-0.21), while nylon has the lowest friction (0.37+/-0.09). PMID:10493141

  2. In vivo virtual intraoperative surgical photoacoustic microscopy

    SciTech Connect

    Han, Seunghoon Kim, Sehui Kim, Jeehyun E-mail: chulhong@postech.edu; Lee, Changho Jeon, Mansik; Kim, Chulhong E-mail: chulhong@postech.edu

    2013-11-11

    We developed a virtual intraoperative surgical photoacoustic microscopy system by combining with a commercial surgical microscope and photoacoustic microscope (PAM). By sharing the common optical path in the microscope and PAM system, we could acquire the PAM and microscope images simultaneously. Moreover, by employing a beam projector to back-project 2D PAM images onto the microscope view plane as augmented reality, the conventional microscopic and 2D cross-sectional PAM images are concurrently mapped on the plane via an ocular lens of the microscope in real-time. Further, we guided needle insertion into phantom ex vivo and mice skins in vivo.

  3. In vivo toxicity study of Lantana camara

    PubMed Central

    Pour, Badakhshan Mahdi; Sasidharan, Sreenivasan

    2011-01-01

    Objective To investigate the toxicity of methanol extract of various parts (Root, Stem, Leaf, Flower and Fruit) of Lantana camara (L. Camara) in Artemia salina. Methods The methanol extracts of L. camara were tested for in vivo brine shrimp lethality assay. Results All the tested extract exhibited very low toxicity on brine shrimp larva. The results showed that the root extract was the most toxic part of L. camara and may have potential as anticancer agent. Conclusions Methanolic extract of L. camara is relatively safe on short-term exposure. PMID:23569765

  4. Imaging Protein-protein Interactions in vivo

    PubMed Central

    Seegar, Tom; Barton, William

    2010-01-01

    Protein-protein interactions are a hallmark of all essential cellular processes. However, many of these interactions are transient, or energetically weak, preventing their identification and analysis through traditional biochemical methods such as co-immunoprecipitation. In this regard, the genetically encodable fluorescent proteins (GFP, RFP, etc.) and their associated overlapping fluorescence spectrum have revolutionized our ability to monitor weak interactions in vivo using Förster resonance energy transfer (FRET)1-3. Here, we detail our use of a FRET-based proximity assay for monitoring receptor-receptor interactions on the endothelial cell surface. PMID:20972411

  5. Laser microspectrofluorometry of photopigments in vivo

    NASA Astrophysics Data System (ADS)

    Colombetti, G.; Ghetti, F.; Lenci, F.; Polacco, E.; Posudin, Yu I.; Campani, E.

    1981-12-01

    A study of the spectral properties of photopigments of microorganisms is of major importance for the understanding of molecular mechanisms whereby these can respond to changes in external illumination conditions. Microspectroscopy in vivo using a tunable dye laser as an excitation source was employed to solve this problem for the case of the unicellular algae Euglena gracilis. This experimental approach made it possible to study fluorescence excitation spectra of photopigments, their average lifetime, and any photochemical reactions which may accompany the absorption of light.

  6. Activities of Psilostachyin A and Cynaropicrin against Trypanosoma cruzi In Vitro and In Vivo

    PubMed Central

    da Silva, Cristiane França; Batista, Denise da Gama Jaen; De Araújo, Julianna Siciliano; Batista, Marcos Meuser; Lionel, Jessica; de Souza, Elen Mello; Hammer, Erica Ripoll; da Silva, Patricia Bernardino; De Mieri, Maria; Adams, Michael; Zimmermann, Stefanie; Hamburger, Matthias; Brun, Reto; Schühly, Wolfgang

    2013-01-01

    In vitro and in vivo activities against Trypanosoma cruzi were evaluated for two sesquiterpene lactones: psilostachyin A and cynaropicrin. Cynaropicrin had previously been shown to potently inhibit African trypanosomes in vivo, and psilostachyin A had been reported to show in vivo effects against T. cruzi, albeit in another test design. In vitro data showed that cynaropicrin was more effective than psilostachyin A. Ultrastructural alterations induced by cynaropicrin included shedding events, detachment of large portions of the plasma membrane, and vesicular bodies and large vacuoles containing membranous structures, suggestive of parasite autophagy. Acute toxicity studies showed that one of two mice died at a cynaropicrin dose of 400 mg/kg of body weight given intraperitoneally (i.p.). Although no major plasma biochemical alterations could be detected, histopathology demonstrated that the liver was the most affected organ in cynaropicrin-treated animals. Although cynaropicrin was as effective as benznidazole against trypomastigotes in vitro, the treatment (once or twice a day) of T. cruzi-infected mice (up to 50 mg/kg/day cynaropicrin) did not suppress parasitemia or protect against mortality induced by the Y and Colombiana strains. Psilostachyin A (0.5 to 50 mg/kg/day given once a day) was not effective in the acute model of T. cruzi infection (Y strain), reaching 100% animal mortality. Our data demonstrate that although it is very promising against African trypanosomes, cynaropicrin does not show efficacy compared to benznidazole in acute mouse models of T. cruzi infection. PMID:23939901

  7. Methods of assessment of thrombosis in vivo

    SciTech Connect

    Dewanjee, M.K.

    1987-01-01

    The contributions of platelets and clotting factors in thrombosis on injured vessel and cardiovascular prostheses have been quantified with several tracers. Thrombus formation in vivo could be measured semiquantitatively in animal models and humans with /sup 111/In-labeled platelets, /sup 123/I- and /sup 131/I-labeled fibrinogen, /sup 111/In-labeled antibody to the fibrinogen receptor on the platelet membrane and to fibrin. Thrombus localization by imaging was possible for large thrombus in vessel with deep injury of thrombogenic surface in the acute phase. A single layer of adherent platelets could not be imaged, due to the high background radioactivity present in blood. Thrombogenicity of grafts was compared with that of contralateral vessel. The dynamic process of platelet deposition could be followed accurately using the in vivo imaging technique. In addition, in vitro quantification permits determination of platelet and fibrin density and of the number of fibrin monomers per platelet in thrombus. The roles of prostacyclin, thromboxane inhibitors, and nonsteroidal antiinflammatory drugs have also been evaluated in animals models and humans. The tracer techniques thus provide invaluable information about platelet-fibrin deposition, its organization and dissolution, and for development of less thrombogenic surfaces for use in cardiovascular prostheses. 53 references.

  8. NANOSTRUCTURED PROBES FOR IN VIVO GENE DETECTION

    PubMed Central

    Bao, Gang; Santangelo, Phillip; Nitin, Nitin; Rhee, Won Jong

    2010-01-01

    The ability to visualize in real-time the expression dynamics and localization of specific RNAs in vivo offers tremendous opportunities for biological and disease studies including cancer detection. However, quantitative methods such as real-time PCR and DNA microarrays rely on the use of cell lysates thus not able to obtain important spatial and temporal information. Fluorescence proteins and other reporter systems cannot image endogenous RNA in living cells. Fluorescence in situ hybridization (FISH) assays require washing to achieve specificity, therefore can only be used with fixed cells. Here we review the recent development of nanostructured probes for living cell RNA detection, and discuss the biological and engineering issues and challenges of quantifying gene expression in vivo. In particular, we describe methods that use oligonucleotide probes, combined with novel delivery strategies, to image the relative level, localization and dynamics of RNA in live cells. Examples of detecting endogenous mRNAs, as well as imaging their subcellular localization are given to illustrate the biological applications, and issues in probe design, delivery and target accessibility are discussed. The nanostructured probes promise to open new and exciting opportunities in sensitive gene detection for a wide range of biological and medical applications. PMID:22138717

  9. In vivo imaging of zebrafish embryogenesis

    PubMed Central

    Keller, Philipp J.

    2013-01-01

    The zebrafish Danio rerio has emerged as a powerful vertebrate model system that lends itself particularly well to quantitative investigations with live imaging approaches, owing to its exceptionally high optical clarity in embryonic and larval stages. Recent advances in light microscopy technology enable comprehensive analyses of cellular dynamics during zebrafish embryonic development, systematic mapping of gene expression dynamics, quantitative reconstruction of mutant phenotypes and the system-level biophysical study of morphogenesis. Despite these technical breakthroughs, it remains challenging to design and implement experiments for in vivo long-term imaging at high spatio-temporal resolution. This article discusses the fundamental challenges in zebrafish long-term live imaging, provides experimental protocols and highlights key prop1erties and capabilities of advanced fluorescence microscopes. The article focuses in particular on experimental assays based on light sheet-based fluorescence microscopy, an emerging imaging technology that achieves exceptionally high imaging speeds and excellent signal-to-noise ratios, while minimizing light-induced damage to the specimen. This unique combination of capabilities makes light sheet microscopy an indispensable tool for the in vivo long-term imaging of large developing organisms. PMID:23523701

  10. Intracranial nonthermal irreversible electroporation: in vivo analysis.

    PubMed

    Garcia, Paulo A; Rossmeisl, John H; Neal, Robert E; Ellis, Thomas L; Olson, John D; Henao-Guerrero, Natalia; Robertson, John; Davalos, Rafael V

    2010-07-01

    Nonthermal irreversible electroporation (NTIRE) is a new minimally invasive technique to treat cancer. It is unique because of its nonthermal mechanism of tumor ablation. Intracranial NTIRE procedures involve placing electrodes into the targeted area of the brain and delivering a series of short but intense electric pulses. The electric pulses induce irreversible structural changes in cell membranes, leading to cell death. We correlated NTIRE lesion volumes in normal brain tissue with electric field distributions from comprehensive numerical models. The electrical conductivity of brain tissue was extrapolated from the measured in vivo data and the numerical models. Using this, we present results on the electric field threshold necessary to induce NTIRE lesions (495-510 V/cm) in canine brain tissue using 90 50-mus pulses at 4 Hz. Furthermore, this preliminary study provides some of the necessary numerical tools for using NTIRE as a brain cancer treatment. We also computed the electrical conductivity of brain tissue from the in vivo data (0.12-0.30 S/m) and provide guidelines for treatment planning and execution. Knowledge of the dynamic electrical conductivity of the tissue and electric field that correlates to lesion volume is crucial to ensure predictable complete NTIRE treatment while minimizing damage to surrounding healthy tissue. PMID:20668843

  11. In Vivo Bioluminescence Imaging of Intratumoral Bacteria.

    PubMed

    Cronin, Michelle; Akin, Ali R; Francis, Kevin P; Tangney, Mark

    2016-01-01

    This chapter describes the use of whole-body bioluminescent imaging (BLI) for the study of bacterial trafficking in live mice, with an emphasis on the use of bacteria in therapy of cancer. Bacteria present an attractive class of vector for cancer therapy, possessing a natural ability to grow preferentially within tumors following systemic administration. Bacteria engineered to express the lux gene cassette permit BLI detection of the bacteria and tumor sites concurrently. The location and levels of bacteria within tumors over time can be readily examined, visualized in two or three dimensions. The method is applicable to a wide range of bacterial species and tumor xenograft types. This article describes the protocol for analysis of bioluminescent bacteria within subcutaneous tumor-bearing mice. This powerful, and inexpensive, real-time imaging strategy represents an ideal method for the study of bacteria in vivo in the context of cancer research. This protocol outlines the procedure for studying lux-tagged Escherichia coli and Bifidobacterium breve in mice, demonstrating the spatial and temporal readout from 2D and 3D BLI achievable with whole-body in vivo luminescence imaging. PMID:26846803

  12. Continuous measurements of ATP secretion in vivo.

    PubMed

    Smith, J B; Burke, S E; Lefer, A M; Freilich, A

    1984-01-01

    Blood was withdrawn continuously from femoral veins of anesthetized rabbits at a rate of 0.07 ml/min. Sodium citrate was pumped into the blood to prevent coagulation, and luciferin-luciferase reagent was added to permit the continuous detection of extracellular ATP. Subsequently, the red blood cells were lysed and the platelet count was recorded continuously. Injection of platelet activating factor or collagen into rabbit ear veins caused an almost immediate but short-lived increase in extracellular ATP with a simultaneous but more prolonged decrease in the platelet count. Although both the endoperoxide analog 9,11-azo-PGH2 and ADP also decreased the platelet count, little extracellular ATP was detected after the azo-PGH2 and none after ADP. These studies demonstrate that those agents that cause platelet secretion from rabbit platelets in vitro also cause secretion in vivo. The method described should be useful in evaluating the capacity of antithrombotic drugs to modify platelet secretion in vivo. PMID:24277184

  13. Biomedical Applications of Sodium MRI In Vivo

    PubMed Central

    Madelin, Guillaume; Regatte, Ravinder R.

    2013-01-01

    In this article, we present an up-to-date overview of the potential biomedical applications of sodium MRI in vivo. Sodium MRI is a subject of increasing interest in translational imaging research as it can give some direct and quantitative biochemical information on the tissue viability, cell integrity and function, and therefore not only help the diagnosis but also the prognosis of diseases and treatment outcomes. It has already been applied in vivo in most of human tissues, such as brain for stroke or tumor detection and therapeutic response, in breast cancer, in articular cartilage, in muscle and in kidney, and it was shown in some studies that it could provide very useful new information not available through standard proton MRI. However, this technique is still very challenging due to the low detectable sodium signal in biological tissue with MRI and hardware/software limitations of the clinical scanners. The article is divided in three parts: (1) the role of sodium in biological tissues, (2) a short review on sodium magnetic resonance, and (3) a review of some studies on sodium MRI on different organs/diseases to date. PMID:23722972

  14. Multidimensional in vivo hazard assessment using zebrafish.

    PubMed

    Truong, Lisa; Reif, David M; St Mary, Lindsey; Geier, Mitra C; Truong, Hao D; Tanguay, Robert L

    2014-01-01

    There are tens of thousands of man-made chemicals in the environment; the inherent safety of most of these chemicals is not known. Relevant biological platforms and new computational tools are needed to prioritize testing of chemicals with limited human health hazard information. We describe an experimental design for high-throughput characterization of multidimensional in vivo effects with the power to evaluate trends relating to commonly cited chemical predictors. We evaluated all 1060 unique U.S. EPA ToxCast phase 1 and 2 compounds using the embryonic zebrafish and found that 487 induced significant adverse biological responses. The utilization of 18 simultaneously measured endpoints means that the entire system serves as a robust biological sensor for chemical hazard. The experimental design enabled us to describe global patterns of variation across tested compounds, evaluate the concordance of the available in vitro and in vivo phase 1 data with this study, highlight specific mechanisms/value-added/novel biology related to notochord development, and demonstrate that the developmental zebrafish detects adverse responses that would be missed by less comprehensive testing strategies. PMID:24136191

  15. In vivo proton range verification: a review

    NASA Astrophysics Data System (ADS)

    Knopf, Antje-Christin; Lomax, Antony

    2013-08-01

    Protons are an interesting modality for radiotherapy because of their well defined range and favourable depth dose characteristics. On the other hand, these same characteristics lead to added uncertainties in their delivery. This is particularly the case at the distal end of proton dose distributions, where the dose gradient can be extremely steep. In practice however, this gradient is rarely used to spare critical normal tissues due to such worries about its exact position in the patient. Reasons for this uncertainty are inaccuracies and non-uniqueness of the calibration from CT Hounsfield units to proton stopping powers, imaging artefacts (e.g. due to metal implants) and anatomical changes of the patient during treatment. In order to improve the precision of proton therapy therefore, it would be extremely desirable to verify proton range in vivo, either prior to, during, or after therapy. In this review, we describe and compare state-of-the art in vivo proton range verification methods currently being proposed, developed or clinically implemented.

  16. Activity ratios of thorium daughters in vivo

    SciTech Connect

    Toohey, R.E.; Rundo, J.; Sha, J.Y.; Essling, M.A.; Pedersen, J.C.; Slane, J.M.

    1984-01-01

    A computerized method of least squares has been used to analyze the /sup 228/Ac and /sup 212/Pb-/sup 212/Bi and daughter ..gamma..-ray spectra obtained in vivo from 133 former workers at a thorium refinery. In addition, the exhalation rate of /sup 220/Rn was determined for each subject and expressed as pCi of emanating /sup 224/Ra. This value was added to the /sup 212/Pb value determined from the ..gamma..-ray measurements to obtain the total /sup 224/Ra present, and the ratio of /sup 224/Ra to /sup 228/Ac was calculated. Values of the ratio ranged from 0.52 +- 0.32 to 2.1 +- 1.7, with a weighted mean of 0.92 +- 0.17. However, it appears that the ratio observed in a given case is characteristic for that case alone; the computed mean value may not be meaningful. The least squares fitting procedure and the overall calibration of the counting system were validated by measurements of /sup 224/Ra in the lungs of one subject postmortem, compared with results obtained from the same subject in vivo. 6 references, 5 figures.

  17. In vivo pharmacodynamic imaging of proteasome inhibition.

    PubMed

    Kimbrel, Erin A; Davis, Tina N; Bradner, James E; Kung, Andrew L

    2009-01-01

    Inhibiting the proteolytic activity of the 26S proteasome has been shown to have selective apoptotic effects on cancer cells and to be clinically efficacious in certain malignancies. There is an unmet medical need for additional proteasome inhibitors, and their development will be facilitated by surrogate markers of proteasome function. Toward this end, ectopic fusion of the destruction domain from ornithine decarboxylase (ODC) to reporter proteins is often used for assessing proteasome function. For luciferase-based reporters, we hypothesized that the oxygen-dependent destruction domain (ODD) from hypoxia-inducible factor 1 alpha (HIF-1 alpha) may provide improved sensitivity over luciferase-ODC, owing to its extremely rapid turnover by the proteasome (HIF-1 alpha has a half-life of less than 5 minutes). In the current study, we show that ODD-luciferase affords a greater dynamic range and faster kinetics than luciferase-ODC in sensing proteasome inhibition in vitro. Importantly, ODD-luciferase also serves as an effective in vivo marker of proteasome function in xenograft tumor models, with inhibition being detected by noninvasive imaging within 3 hours of bortezomib administration. These data establish ODD-luciferase as a surrogate marker of proteasome function that can be used both in vitro and in vivo for the development of novel proteasome inhibitors. PMID:19723471

  18. Spectroscopy analysis of tissues in vivo

    NASA Astrophysics Data System (ADS)

    Loschenov, Victor B.; Poleshkin, P. V.; Stratonnikov, Alexander A.; Torshina, Nadezgda L.

    1995-01-01

    The spectral analysis of biological tissues in vivo is widely used in various fields particularly in medical diagnostics and therapy control. Great possibilities of spectral tissue analysis exist to be realized in the future. Among them are the complete non-invasive clinical blood analysis with evaluation of, for example, sugar concentration in blood; the evaluation of chemical state and localization on subcell level of various drugs binded with biological structures. These facts were shown to affect drastically the drug therapeutic activity. The main advantage of spectral analysis of tissues in vivo is its noninvasivity. This allows one to get information about tissue condition without affecting the dynamic of various biological processes. Another advantage of optical tissue analysis is the possibility to process data in real time and to control parameters of therapy process according to information acquired. For example the in situ analysis of photosensitizer concentration and its chemical state during photodynamic therapy makes it possible to correct the laser irradiation intensity (the photobleaching of photosensitizer requires the decrease in laser intensity).

  19. In vivo identification of periodontal progenitor cells.

    PubMed

    Roguljic, H; Matthews, B G; Yang, W; Cvija, H; Mina, M; Kalajzic, I

    2013-08-01

    The periodontal ligament contains progenitor cells; however, their identity and differentiation potential in vivo remain poorly characterized. Previous results have suggested that periodontal tissue progenitors reside in perivascular areas. Therefore, we utilized a lineage-tracing approach to identify and track periodontal progenitor cells from the perivascular region in vivo. We used an alpha-smooth muscle actin (αSMA) promoter-driven and tamoxifen-inducible Cre system (αSMACreERT2) that, in combination with a reporter mouse line (Ai9), permanently labels a cell population, termed 'SMA9'. To trace the differentiation of SMA9-labeled cells into osteoblasts/cementoblasts, we utilized a Col2.3GFP transgene, while expression of Scleraxis-GFP was used to follow differentiation into periodontal ligament fibroblasts during normal tissue formation and remodeling following injury. In uninjured three-week-old SMA9 mice, tamoxifen labeled a small population of cells in the periodontal ligament that expanded over time, particularly in the apical region of the root. By 17 days and 7 weeks after labeling, some SMA9-labeled cells expressed markers indicating differentiation into mature lineages, including cementocytes. Following injury, SMA9 cells expanded, and differentiated into cementoblasts, osteoblasts, and periodontal ligament fibroblasts. SMA9-labeled cells represent a source of progenitors that can give rise to mature osteoblasts, cementoblasts, and fibroblasts within the periodontium. PMID:23735585

  20. Towards Quantitative Phosphotyrosine Profiling In Vivo

    PubMed Central

    Johnson, Hannah; White, Forest M.

    2012-01-01

    Tyrosine phosphorylation is a dynamic reversible post-translational modification that regulates many aspects of cell biology. To understand how this modification controls biological function, it is necessary to not only identify the specific sites of phosphorylation, but also to quantify how phosphorylation levels on these sites may be altered under specific physiological conditions. Due to its sensitivity and accuracy, mass spectrometry (MS) has widely been applied to the identification and characterization of phosphotyrosine signaling across biological systems. In this review we highlight the advances in both MS and phosphotyrosine enrichment methods that have been developed to enable the identification of low level tyrosine phosphorylation events. Computational and manual approaches to ensure confident identification of phosphopeptide sequence and determination of phosphorylation site localization are discussed along with methods that have been applied to the relative quantification of large numbers of phosphorylation sites. Finally, we provide an overview of the challenges ahead as we extend these technologies to the characterization of tyrosine phosphorylation signaling in vivo. With these latest developments in analytical and computational techniques, it is now possible to derive biological insight from quantitative MS-based analysis of signaling networks in vitro and in vivo. Application of these approaches to a wide variety of biological systems will define how signal transduction regulates cellular physiology in health and disease. PMID:22677333

  1. Nutritional abrogation of photoimmunosuppression: in vivo investigations.

    PubMed

    Pilkington, Suzanne M; Gibbs, Neil K; Friedmann, Peter S; Rhodes, Lesley E

    2014-01-01

    Skin cancer is a major public health concern, and the primary aetiological factor in the majority of skin cancers is ultraviolet radiation (UVR) exposure. UVR not only induces potentially mutagenic DNA damage but also suppresses cell-mediated immunity (CMI), allowing cancerous cells to escape destruction and progress to tumours. A considerable proportion of an individual's annual sun exposure is obtained outside the vacation period when topical and physical measures for photoprotection are irregularly used. Certain nutrients could provide an adjunctive protective role, and evidence is accruing from experimental studies to support their use in abrogation of photoimmunosuppression. Moreover, developments in clinical research methods to evaluate impact of solar-simulated radiation on cutaneous CMI allow the immune protective potential of nutritional agents to be examined in humans in vivo. This article summarises the mediation of CMI and its suppression by UVR, evaluates the methodology for quantitative assessment in vivo, reviews the human studies reported on nutritional abrogation of photoimmunosuppression including recent randomized controlled trials and discusses the mechanisms of photoprotection by the nutrients. This includes, in addition to antioxidants, novel studies of omega-3 polyunsaturated fatty acids and nicotinamide. PMID:24283330

  2. Netrin-4 induces lymphangiogenesis in vivo.

    PubMed

    Larrieu-Lahargue, Frederic; Welm, Alana L; Thomas, Kirk R; Li, Dean Y

    2010-07-01

    Netrin-4, a laminin-related secreted protein is an axon guidance cue recently shown essential outside of the nervous system, regulating mammary and lung morphogenesis as well as blood vascular development. Here, we show that Netrin-4, at physiologic doses, induces proliferation, migration, adhesion, tube formation and survival of human lymphatic endothelial cells in vitro comparable to well-characterized lymphangiogenic factors fibroblast growth factor-2 (FGF-2), hepatocyte growth factor (HGF), vascular endothelial growth factor-A (VEGF-A), and vascular endothelial growth factor-C (VEGF-C). Netrin-4 stimulates phosphorylation of intracellular signaling components Akt, Erk and S6, and their specific inhibition antagonizes Netrin-4-induced proliferation. Although Netrin receptors Unc5B and neogenin, are expressed by human lymphatic endothelial cells, suppression of either or both does not suppress Netrin-4-promoted in vitro effects. In vivo, Netrin-4 induces growth of lymphatic and blood vessels in the skin of transgenic mice and in breast tumors. Its overexpression in human and mouse mammary carcinoma cancer cells leads to enhanced metastasis. Finally, Netrin-4 stimulates in vitro and in vivo lymphatic permeability by activating small GTPases and Src family kinases/FAK, and down-regulating tight junction proteins. Together, these data provide evidence that Netrin-4 is a lymphangiogenic factor contributing to tumor dissemination and represents a potential target to inhibit metastasis formation. PMID:20407033

  3. Aggregation states of phosphoribulokinase (PRK) in vivo

    SciTech Connect

    Porter, M.A.; Hartman, F.C. )

    1989-04-01

    Spinach PRK, extracted from either light- or dark-harvested tissue (LHT or DHT) in the presence of DTT, has a M{sub r} of 90 kDa and is fully active. Consistent with an earlier study extraction of LHT in the absence of DTT results in two forms of inactive PRK, M{sub r} 90 kDa (LMW) and M{sub r}> 550 kDa (HMW). If 400 mM (NH{sub 4}){sub 2}SO{sub 4} without DTT is included during extraction, the active LMW predominates implicating it as the major, functional form in vivo during periods of illumination. Either high- or low-sale extraction of DHT reveals mostly HMW; prolonged incubation of the high-salt extract causes disaggregation of LMW without activation. These data suggest that the dark form of PRK in vivo is an aggregate, formed by either self-association or by interactions with other proteins. Salt-induced disaggregation of HMW is inconsistent with intermolecular disulfides crosslinking the aggregated PRK; therefore, oxidation-induced conformational changes must promote aggregation.

  4. Aneuploidy in mammalian somatic cells in vivo.

    PubMed

    Cimino, M C; Tice, R R; Liang, J C

    1986-01-01

    Aneuploidy is an important potential source of human disease and of reproductive failure. Nevertheless, the ability of chemical agents to induce aneuploidy has been investigated only sporadically in intact (whole-animal) mammalian systems. A search of the available literature from the EMCT Aneuploidy File (for years 1970-1983) provided 112 papers that dealt with aneuploidy in mammalian somatic cells in vivo. 59 of these papers did not meet minimal criteria for analysis and were rejected from subsequent review. Of the remaining 53 papers that dealt with aneuploidy induction by chemical agents in mammalian somatic cells in vivo, only 3 (6%) contained data that were considered to be supported conclusively by adequate study designs, execution, and reporting. These 3 papers dealt with 2 chemicals, one of which, mercury, was negative for aneuploidy induction in humans, and the other, pyrimethamine, was positive in an experimental rodent study. The majority of papers (94%) were considered inconclusive for a variety of reasons. The most common reasons for calling a study inconclusive were (a) combining data on hyperploidy with those on hypoploidy and/or polyploidy, (b) an inadequate or unspecified number of animals and/or cells per animal scored per treatment group, and (c) poor data presentation such that animal-to-animal variability could not be assessed. Suggestions for protocol development are made, and the future directions of research into aneuploidy induction are discussed. PMID:3941670

  5. Multidimensional In Vivo Hazard Assessment Using Zebrafish

    PubMed Central

    Tanguay, Robert L.

    2014-01-01

    There are tens of thousands of man-made chemicals in the environment; the inherent safety of most of these chemicals is not known. Relevant biological platforms and new computational tools are needed to prioritize testing of chemicals with limited human health hazard information. We describe an experimental design for high-throughput characterization of multidimensional in vivo effects with the power to evaluate trends relating to commonly cited chemical predictors. We evaluated all 1060 unique U.S. EPA ToxCast phase 1 and 2 compounds using the embryonic zebrafish and found that 487 induced significant adverse biological responses. The utilization of 18 simultaneously measured endpoints means that the entire system serves as a robust biological sensor for chemical hazard. The experimental design enabled us to describe global patterns of variation across tested compounds, evaluate the concordance of the available in vitro and in vivo phase 1 data with this study, highlight specific mechanisms/value-added/novel biology related to notochord development, and demonstrate that the developmental zebrafish detects adverse responses that would be missed by less comprehensive testing strategies. PMID:24136191

  6. In vivo imaging of neuroinflammation in schizophrenia.

    PubMed

    Pasternak, Ofer; Kubicki, Marek; Shenton, Martha E

    2016-06-01

    In recent years evidence has accumulated to suggest that neuroinflammation might be an early pathology of schizophrenia that later leads to neurodegeneration, yet the exact role in the etiology, as well as the source of neuroinflammation, are still not known. The hypothesis of neuroinflammation involvement in schizophrenia is quickly gaining popularity, and thus it is imperative that we have reliable and reproducible tools and measures that are both sensitive, and, most importantly, specific to neuroinflammation. The development and use of appropriate human in vivo imaging methods can help in our understanding of the location and extent of neuroinflammation in different stages of the disorder, its natural time-course, and its relation to neurodegeneration. Thus far, there is little in vivo evidence derived from neuroimaging methods. This is likely the case because the methods that are specific and sensitive to neuroinflammation are relatively new or only just being developed. This paper provides a methodological review of both existing and emerging positron emission tomography and magnetic resonance imaging techniques that identify and characterize neuroinflammation. We describe \\how these methods have been used in schizophrenia research. We also outline the shortcomings of existing methods, and we highlight promising future techniques that will likely improve state-of-the-art neuroimaging as a more refined approach for investigating neuroinflammation in schizophrenia. PMID:26048294

  7. In vivo peripheral nervous system insulin signaling

    PubMed Central

    Grote, Caleb W.; Ryals, Janelle M.; Wright, Douglas E.

    2014-01-01

    Alterations in peripheral nervous system (PNS) insulin support may contribute to diabetic neuropathy (DN); yet, PNS insulin signaling is not fully defined. Here, we investigated in vivo insulin signaling in the PNS and compared the insulin-responsiveness to that of muscle, liver, and adipose. Nondiabetic mice were administered increasing doses of insulin to define a dose response relationship between insulin and Akt activation in the DRG and sciatic nerve. Resulting EC50 doses were used to characterize the PNS insulin signaling time course and make comparisons between insulin signaling in the PNS and other peripheral tissues (i.e., muscle, liver, adipose). The results demonstrate that the PNS is responsive to insulin and that differences in insulin signaling pathway activation exist between PNS compartments. At a therapeutically relevant dose, Akt was activated in the muscle, liver, and adipose at 30 minutes, correlating with the changes in blood glucose levels. Interestingly, the sciatic nerve showed a similar signaling profile as insulin-sensitive tissues, however there was not a comparable activation in the DRG or spinal cord. These results present new evidence regarding PNS insulin signaling pathways in vivo and provide a baseline for studies investigating the contribution of disrupted PNS insulin signaling to DN pathogenesis. PMID:24028189

  8. Use of RSP Tooling to Manufacture Die Casting Dies

    SciTech Connect

    Kevin McHugh

    2004-07-01

    The technology and art used to construct die casting dies has seen many improvements over the years. However, the time lag from when a design is finalized to the time a tool is in production has remained essentially the same. The two main causes for the bottleneck are the need to qualify a part design by making prototypes (usually from an alternative process), and the production tooling lead time after the prototypes are approved. Production tooling costs are high due to the labor and equipment costs associated with transforming a forged block of tool steel into a finished tool. CNC machining, sink EDM, benching, engraving and heat treatment unit operations are typically involved. As a result, there is increasing interest in rapid tooling (RT) technologies that shorten the design-to-part cycle and reduce the cost of dies. There are currently more than 20 RT methods being developed and refined around the world (1). The "rapid" in rapid tooling suggests time compression for tool delivery, but does not address robustness as nearly all RT approaches are intended for low-volume prototype work, primarily for molding plastics. Few options exist for die casting. An RT technology suitable for production-quality tooling in the time it normally takes for prototype tooling is highly desirable. In fact, there would be no need for a distinction between prototype and production tooling. True prototype parts could be made using the same processing conditions and materials intended for production. Qualification of the prototype part would allow the manufacturer to go directly into production with the same tool. A relatively new RT technology, Rapid Solidification Process (RSP) Tooling, is capable of making production-quality tooling in an RT timeframe for die casting applications. RSP Tooling, was developed at the Idaho National Engineering and Environmental Laboratory (INEEL), and commercialized with the formation of RSP Tooling, LLC (2). This paper describes the process, and

  9. Blood rheology in vitro and in vivo.

    PubMed

    Lowe, G D

    1987-09-01

    Blood rheology tests are traditionally used for detection of organic disease and for monitoring disease activity. More recently they have been used for prediction of blood flow in vivo, not only in overt hyperviscosity syndromes but also in the covert hyperviscosity of low-flow states. The traditional ESR test result increases with red cell aggregation induced by increases in large, asymmetrical plasma globulins. However, small increases in haematocrit and large increases in plasma viscosity each decrease the ESR, reducing both its diagnostic utility and its ability to predict blood flow in vivo. The ESR should be corrected to a standard haematocrit, or else replaced by the ZSR or plasma viscosity, which are more rapid, simple, sensitive and independent of haematocrit. For prediction of blood flow in vivo, these tests can be supplemented by measurement of whole-blood viscosity, which can be performed simply and cheaply in capillary viscometers at high shear rates. Whole-blood viscosity is determined by plasma viscosity, haematocrit and red cell deformability at high shear rates. Its measurement is useful in overt hyperviscosity syndromes, particularly in estimating the effect of red cell transfusion in anaemic patients with plasma hyperviscosity, hyperleukocytic leukaemias or sickling disorders. Blood viscosity should be related to the haematocrit or haemoglobin concentration in order to estimate oxygen delivery to tissues. Changes in blood viscosity can be compensated readily in the normal circulation but not in the compromised, low-flow circulation. In these circumstances, systemic increases in plasma viscosity, haematocrit, whole-blood viscosity, red cell aggregation and in the numbers of circulating rigid red or white blood cells can perpetuate low-flow states and ischaemia. Red cell deformability in narrow vessels is best measured by micropore filtration systems, in which the effect of white cells has been eliminated. Red cell deformability is reduced by

  10. Acting to let someone die.

    PubMed

    McGee, Andrew

    2015-02-01

    This paper examines the recent prominent view in medical ethics that withdrawing life-sustaining treatment (LST) is an act of killing. I trace this view to the rejection of the traditional claim that withdrawing LST is an omission rather than an act. Although that traditional claim is not as problematic as this recent prominent view suggests, my main claim is that even if we accepted that withdrawing LST should be classified as an act rather than as an omission, it could still be classified as letting die rather than killing. Even though omissions are contrasted with acts, letting die need not be, for one can let die by means of acts. The remainder of the paper is devoted to establishing this claim and addresses certain objections to it. PMID:24320715

  11. Clinical management of dying patients.

    PubMed Central

    Gavrin, J; Chapman, C R

    1995-01-01

    Dying is universal, and death should be a peaceful time. Myriad comfort measures are available in the last weeks before life ends. Discussions about end-of-life issues often suffer from lack of informed opinion. Palliative care experts have identified specific somatic and psychological sources of distress for dying patients and their loved ones. Pain, shortness of breath, nausea and vomiting, and fear of abandonment contribute substantially to both physical and psychological discomfort toward the end of life. Simple, effective methods exist for relieving those symptoms. Knowledge about the natural events associated with dying and an informed approach to medical and psychological interventions contribute to systematic and successful comfort care. We describe the origin of physical and psychological distress at the end of life and provide strategies for alleviating many of the discomforts. PMID:7571591

  12. 'Die Zeit' im Konversationsunterricht ('Die Zeit' in a Conversation Class)

    ERIC Educational Resources Information Center

    Becker-Cantarino, Barbel

    1975-01-01

    The German weekly newspaper "Die Zeit" contains a very rich variety of topics and is therefore an inexhaustable source for an exciting conversation class. This article outlines and lists the advantages of a German conversation course organized around this newspaper. (Text is in German.) (TL)

  13. Portable punch and die jig

    DOEpatents

    Lewandowski, Edward F.; Anderson, Petrus A.

    1978-01-01

    A portable punch and die jig includes a U-shaped jig of predetermined width having a slot of predetermined width in the base thereof extending completely across the width of the jig adapted to fit over the walls of rectangular tubes and a punch and die assembly disposed in a hole extending through the base of the jig communicating with the slot in the base of the jig for punching a hole in the walls of the rectangular tubes at precisely determined locations.

  14. Dying radio galaxies in clusters

    NASA Astrophysics Data System (ADS)

    Murgia, M.; Parma, P.; Mack, K.-H.; de Ruiter, H. R.; Fanti, R.; Govoni, F.; Tarchi, A.; Giacintucci, S.; Markevitch, M.

    2011-02-01

    Aims: We present a study of five "dying" nearby (z ≤ 0.2) radio galaxies belonging to both the WENSS minisurvey and the B2 bright catalogs WNB1734+6407, WNB1829+6911, WNB1851+5707, B2 0120+33, and B2 1610+29. Methods: These sources have been selected on the basis of their extremely steep broad-band radio spectra, which strongly indicates that either these objects belong to the rare class of dying radio galaxies or we are observing "fossil" radio plasma remaining from a previous instance of nuclear activity. We derive the relative duration of the dying phase from the fit of a synchrotron radiative model to the radio spectra of the sources. Results: The modeling of the integrated spectra and the deep spectral index images obtained with the VLA confirmed that in these sources the central engine has ceased to be active for a significant fraction of their lifetime, although their extended lobes have not yet completely faded away. We found that WNB1851+5707 is in reality composed of two distinct dying galaxies, which appear blended together as a single source in the WENSS. In the cases of WNB1829+6911 and B2 0120+33, the fossil radio lobes are seen in conjunction with a currently active core. A very faint core is also detected in a MERLIN image of WNB1851+5707a, one of the two dying sources composing WNB1851+5707. We found that all sources in our sample are located (at least in projection) at the center of an X-ray emitting cluster. Conclusions: Our results suggest that the duration of the dying phase for a radio source in a cluster can be significantly higher than that of a radio galaxy in the field, although no firm conclusions can be drawn because of the small number statistics involved. The simplest interpretation of the tendency for dying galaxies to be found in clusters is that the low-frequency radio emission from the fading radio lobes lasts longer if their expansion is somewhat reduced or even stopped. Another possibility is that the occurrence of dying

  15. Die Herz-Lungen-Maschine

    NASA Astrophysics Data System (ADS)

    Krane, Markus; Bauernschmitt, Robert; Lange, Rüdiger

    Das Kapitel der modernen Herzchirurgie mit Einsatz der Herz-Lungen-Maschine am Menschen beginnt am 6. Mai 1953, als J. Gibbon bei einer 18-jährigen Patientin einen angeborenen Defekt in der Vorhofscheidewand verschließt [1]. Mit ersten experimentellen Versuchen zur extrakorporalen Zirkulation begann Gibbon bereits in den 30er Jahren des 20. Jahrhunderts. Die Grundlage für die heute gebräuchliche Rollerpumpe schufen Porter und Bradley mit ihrer "rotary pump“, welche sie 1855 zum Patent anmeldeten. Diese Pumpe wurde von DeBakey und Schmidt modifiziert und entspricht im Wesentlichen noch der heute sich im Routinebetrieb befindlichen Rollerpumpe [2].

  16. Nucleosome dynamics during chromatin remodeling in vivo

    PubMed Central

    Ramachandran, Srinivas; Henikoff, Steven

    2016-01-01

    ABSTRACT Precise positioning of nucleosomes around regulatory sites is achieved by the action of chromatin remodelers, which use the energy of ATP to slide, evict or change the composition of nucleosomes. Chromatin remodelers act to bind nucleosomes, disrupt histone-DNA interactions and translocate the DNA around the histone core to reposition nucleosomes. Hence, remodeling is expected to involve nucleosomal intermediates with a structural organization that is distinct from intact nucleosomes. We describe the identification of a partially unwrapped nucleosome structure using methods that map histone-DNA contacts genome-wide. This alternative nucleosome structure is likely formed as an intermediate or by-product during nucleosome remodeling by the RSC complex. Identification of the loss of histone-DNA contacts during chromatin remodeling by RSC in vivo has implications for the regulation of transcriptional initiation. PMID:26933790

  17. Corneal In Vivo Confocal Microscopy: Clinical Applications.

    PubMed

    You, Jae Young; Botelho, Paul J

    2016-01-01

    In vivo confocal microscopy (IVCM) has become a widely accepted imaging technique to study the human living cornea. It provides a unique opportunity to visualize the corneal tissue at the cellular level without damage and longitudinally observe its pathologic and normative changes. With rapidly evolving technology, there has been an abundance of interest in maximizing its potential to better understand the human cornea in health and disease. This is evidenced by a growing literature analyzing acquired and inherited corneal and also systemic diseases using corneal IVCM. This article provides a narrative review of IVCM and its applications. [Full article available at http://rimed.org/rimedicaljournal-2016-06.asp, free with no login]. PMID:27247970

  18. Modeling Melanoma In Vitro and In Vivo

    PubMed Central

    Beaumont, Kimberley A.; Mohana-Kumaran, Nethia; Haass, Nikolas K.

    2013-01-01

    The behavior of melanoma cells has traditionally been studied in vitro in two-dimensional cell culture with cells adhering to plastic dishes. However, in order to mimic the three-dimensional architecture of a melanoma, as well as its interactions with the tumor microenvironment, there has been the need for more physiologically relevant models. This has been achieved by designing 3D in vitro models of melanoma, such as melanoma spheroids embedded in extracellular matrix or organotypic skin reconstructs. In vivo melanoma models have typically relied on the growth of tumor xenografts in immunocompromised mice. Several genetically engineered mouse models have now been developed which allow the generation of spontaneous melanoma. Melanoma models have also been established in other species such as zebrafish, which are more conducive to imaging and high throughput studies. We will discuss these models as well as novel techniques that are relevant to the study of the molecular mechanisms underlying melanoma progression.

  19. In Vivo Molecular Imaging in Retinal Disease

    PubMed Central

    Xie, Fang; Luo, Wenting; Zhang, Zhongyu; Sun, Dawei

    2012-01-01

    There is an urgent need for early diagnosis in medicine, whereupon effective treatments could prevent irreversible tissue damage. The special structure of the eye provides a unique opportunity for noninvasive light-based imaging of ocular fundus vasculature. To detect endothelial injury at the early and reversible stage of adhesion molecule upregulation, some novel imaging agents that target retinal endothelial molecules were generated. In vivo molecular imaging has a great potential to impact medicine by detecting diseases or screening disease in early stages, identifying extent of disease, selecting disease and patient-specific therapeutic treatment, applying a directed or targeted therapy, and measuring molecular-specific effects of treatment. Current preclinical findings and advances in instrumentation such as endoscopes and microcatheters suggest that these molecular imaging modalities have numerous clinical applications and will be translated into clinical use in the near future. PMID:22363836

  20. Spectral reflectance of human skin in vivo.

    PubMed

    Andersen, P H; Bjerring, P

    1990-02-01

    A newly developed skin reflectance spectrophotometer was evaluated for measurements of both melanin pigmentation and erythema. Physiological changes in blood flow and blood content in normal humans were induced by compression with an arm cuff during recording of skin reflectance spectra. Reflectance spectra of UV-induced erythema were also recorded and compared with laser-Doppler flow measurements. Spectral reflectance measurements were found to be highly sensitive in determining minimal erythema, which was not clinically detectable. The measurements of erythema using reflectance spectroscopy and UV irradiation were very highly correlated (r = 0.996). It was possible to calculate the in vivo absorbance of oxygenized haemoglobin. The melanin pigmentation following UV irradiation was quantified by reflectance spectroscopy and correlates highly with the dose of UV irradiation (r = 0.995). Furthermore, regional variations in skin melanin and haemoglobin were analysed for fair Caucasian skin. PMID:2196543

  1. In Vivo Cytogenetic Studies on Aspartame

    PubMed Central

    AlSuhaibani, Entissar S.

    2010-01-01

    Aspartame (a-Laspartyl-L-phenylalanine 1-methylester) is a dipeptide low-calorie artificial sweetener that is widely used as a nonnutritive sweetener in foods and drinks. The safety of aspartame and its metabolic breakdown products (phenylalanine, aspartic acid and methanol) was investigated in vivo using chromosomal aberration (CA) test and sister chromatid exchange (SCE) test in the bone marrow cells of mice. Swiss Albino male mice were exposed to aspartame (3.5, 35, 350 mg/kg body weight). Bone marrow cells isolated from femora were analyzed for chromosome aberrations and sister chromatid exchanges. Treatment with aspartame induced dose dependently chromosome aberrations at all concentrations while it did not induce sister chromatid exchanges. On the other hand, aspartame did not decrease the mitotic index (MI). However, statistical analysis of the results show that aspartame is not significantly genotoxic at low concentration. PMID:20689731

  2. THz Medical Imaging: in vivo Hydration Sensing

    PubMed Central

    Taylor, Zachary D.; Singh, Rahul S.; Bennett, David B.; Tewari, Priyamvada; Kealey, Colin P.; Bajwa, Neha; Culjat, Martin O.; Stojadinovic, Alexander; Lee, Hua; Hubschman, Jean-Pierre; Brown, Elliott R.; Grundfest, Warren S.

    2015-01-01

    The application of THz to medical imaging is experiencing a surge in both interest and federal funding. A brief overview of the field is provided along with promising and emerging applications and ongoing research. THz imaging phenomenology is discussed and tradeoffs are identified. A THz medical imaging system, operating at ~525 GHz center frequency with ~125 GHz of response normalized bandwidth is introduced and details regarding principles of operation are provided. Two promising medical applications of THz imaging are presented: skin burns and cornea. For burns, images of second degree, partial thickness burns were obtained in rat models in vivo over an 8 hour period. These images clearly show the formation and progression of edema in and around the burn wound area. For cornea, experimental data measuring the hydration of ex vivo porcine cornea under drying is presented demonstrating utility in ophthalmologic applications. PMID:26085958

  3. Quantifying drug-protein binding in vivo.

    SciTech Connect

    Buchholz, B; Bench, G; Keating III, G; Palmblad, M; Vogel, J; Grant, P G; Hillegonds, D

    2004-02-17

    Accelerator mass spectrometry (AMS) provides precise quantitation of isotope labeled compounds that are bound to biological macromolecules such as DNA or proteins. The sensitivity is high enough to allow for sub-pharmacological (''micro-'') dosing to determine macromolecular targets without inducing toxicities or altering the system under study, whether it is healthy or diseased. We demonstrated an application of AMS in quantifying the physiologic effects of one dosed chemical compound upon the binding level of another compound in vivo at sub-toxic doses [4].We are using tissues left from this study to develop protocols for quantifying specific binding to isolated and identified proteins. We also developed a new technique to quantify nanogram to milligram amounts of isolated protein at precisions that are comparable to those for quantifying the bound compound by AMS.

  4. Seeing Stem Cells at Work In Vivo

    PubMed Central

    Srivastava, Amit K.; Bulte, Jeff W. M.

    2013-01-01

    Stem cell based-therapies are novel therapeutic strategies that hold key for developing new treatments for diseases conditions with very few or no cures. Although there has been an increase in the number of clinical trials involving stem cell-based therapies in the last few years, the long-term risks and benefits of these therapies are still unknown. Detailed in vivo studies are needed to monitor the fate of transplanted cells, including their distribution, differentiation, and longevity over time. Advancements in non-invasive cellular imaging techniques to track engrafted cells in real-time present a powerful tool for determining the efficacy of stem cell-based therapies. In this review, we describe the latest approaches to stem cell labeling and tracking using different imaging modalities. PMID:23975604

  5. An excitatory GABA loop operating in vivo

    PubMed Central

    Astorga, Guadalupe; Bao, Jin; Marty, Alain; Augustine, George J.; Franconville, Romain; Jalil, Abdelali; Bradley, Jonathan; Llano, Isabel

    2015-01-01

    While it has been proposed that the conventional inhibitory neurotransmitter GABA can be excitatory in the mammalian brain, much remains to be learned concerning the circumstances and the cellular mechanisms governing potential excitatory GABA action. Using a combination of optogenetics and two-photon calcium imaging in vivo, we find that activation of chloride-permeable GABAA receptors in parallel fibers (PFs) of the cerebellar molecular layer of adult mice causes parallel fiber excitation. Stimulation of PFs at submaximal stimulus intensities leads to GABA release from molecular layer interneurons (MLIs), thus creating a positive feedback loop that enhances excitation near the center of an activated PF bundle. Our results imply that elevated chloride concentration can occur in specific intracellular compartments of mature mammalian neurons and suggest an excitatory role for GABAA receptors in the cerebellar cortex of adult mice. PMID:26236197

  6. Nucleosome dynamics during chromatin remodeling in vivo.

    PubMed

    Ramachandran, Srinivas; Henikoff, Steven

    2016-01-01

    Precise positioning of nucleosomes around regulatory sites is achieved by the action of chromatin remodelers, which use the energy of ATP to slide, evict or change the composition of nucleosomes. Chromatin remodelers act to bind nucleosomes, disrupt histone-DNA interactions and translocate the DNA around the histone core to reposition nucleosomes. Hence, remodeling is expected to involve nucleosomal intermediates with a structural organization that is distinct from intact nucleosomes. We describe the identification of a partially unwrapped nucleosome structure using methods that map histone-DNA contacts genome-wide. This alternative nucleosome structure is likely formed as an intermediate or by-product during nucleosome remodeling by the RSC complex. Identification of the loss of histone-DNA contacts during chromatin remodeling by RSC in vivo has implications for the regulation of transcriptional initiation. PMID:26933790

  7. In Vivo Dedifferentiation of Adult Adipose Cells

    PubMed Central

    Lu, Feng; Dong, Ziqing; Chang, Qiang; Gao, Jianhua

    2015-01-01

    Introduction Adipocytes can dedifferentiate into fibroblast-like cells in vitro and thereby acquire proliferation and multipotent capacities to participate in the repair of various organs and tissues. Whether dedifferentiation occurs under physiological or pathological conditions in vivo is unknown. Methods A tissue expander was placed under the inguinal fat pads of rats and gradually expanded by injection of water. Samples were collected at various time points, and morphological, histological, cytological, ultrastructural, and gene expression analyses were conducted. In a separate experiment, purified green fluorescent protein+ adipocytes were transplanted into C57 mice and collected at various time points. The transplanted adipocytes were assessed by bioluminescence imaging and whole-mount staining. Results The expanded fat pad was obviously thinner than the untreated fat pad on the opposite side. It was also tougher in texture and with more blood vessels attached. Hematoxylin and eosin staining and transmission electron microscopy indicated there were fewer monolocular adipocytes in the expanded fat pad and the morphology of these cells was altered, most notably their lipid content was discarded. Immunohistochemistry showed that the expanded fat pad contained an increased number of proliferative cells, which may have been derived from adipocytes. Following removal of the tissue expander, many small adipocytes were observed. Bioluminescence imaging suggested that some adipocytes survived when transplanted into an ischemic-hypoxic environment. Whole-mount staining revealed that surviving adipocytes underwent a process similar to adipocyte dedifferentiation in vitro. Monolocular adipocytes became multilocular adipocytes and then fibroblast-like cells. Conclusions Mature adipocytes may be able to dedifferentiate in vivo, and this may be an adipose tissue self-repair mechanism. The capacity of adipocytes to dedifferentiate into stem cell-like cells may also have a

  8. In vivo nanotoxicity assays in plant models.

    PubMed

    Kumari, Mamta; Ernest, Vinita; Mukherjee, Amitava; Chandrasekaran, Natarajan

    2012-01-01

    Increasing application of silver nanoparticles (SNPs) and zinc oxide nanoparticles (nZnO) in consumer products like textiles, cosmetics, washing machines and other household products increases their chance to reach the environment. Intensive research is required to assess the nanoparticles' toxicity to the environmental system. The toxicological effect of nanoparticles has been studied at the miniscule scale and requires intensive research to be conducted to assess its unknown effects. Plants are the primary target species which need to be included to develop a comprehensive toxicity profile for nanoparticles. So far, the mechanisms of toxicity of nanoparticles to the plant system remains largely unknown and little information on the potential uptake of nanoparticles by plants and their subsequent fate within the food chain is available. The phytoxicological behaviour of silver and zinc oxide nanoparticles on Allium cepa and seeds of Zea mays (maize), Cucumis sativus (cucumber) and Lycopersicum esculentum (tomato) was done. The in vitro studies on A. cepa have been done to check the cytotoxicological effects including mitotic index, chromosomal aberrations, vagrant chromosomes, sticky chromosomes, disturbed metaphase, breaks and formation of micronucleus. In vitro and in vivo studies on seed systems exposed to different concentration of nanoparticles dispersion to check phytotoxicity end point as root length, germination effect, adsorption and accumulation of nanoparticles (uptake studies) into the plant systems. In vivo studies in a seed system was done using phytagel medium. Biochemical studies were done to check effect on protein, DNA and thiobarbituric acid reactive species concentration. FT-IR studies were done to analyze the functional and conformational changes in the treated and untreated samples. The toxicological effects of nanoparticles had to be studied at the miniscule scale to address existing environment problems or prevent future problems. The

  9. 3D ultrafast ultrasound imaging in vivo

    NASA Astrophysics Data System (ADS)

    Provost, Jean; Papadacci, Clement; Esteban Arango, Juan; Imbault, Marion; Fink, Mathias; Gennisson, Jean-Luc; Tanter, Mickael; Pernot, Mathieu

    2014-10-01

    Very high frame rate ultrasound imaging has recently allowed for the extension of the applications of echography to new fields of study such as the functional imaging of the brain, cardiac electrophysiology, and the quantitative imaging of the intrinsic mechanical properties of tumors, to name a few, non-invasively and in real time. In this study, we present the first implementation of Ultrafast Ultrasound Imaging in 3D based on the use of either diverging or plane waves emanating from a sparse virtual array located behind the probe. It achieves high contrast and resolution while maintaining imaging rates of thousands of volumes per second. A customized portable ultrasound system was developed to sample 1024 independent channels and to drive a 32  ×  32 matrix-array probe. Its ability to track in 3D transient phenomena occurring in the millisecond range within a single ultrafast acquisition was demonstrated for 3D Shear-Wave Imaging, 3D Ultrafast Doppler Imaging, and, finally, 3D Ultrafast combined Tissue and Flow Doppler Imaging. The propagation of shear waves was tracked in a phantom and used to characterize its stiffness. 3D Ultrafast Doppler was used to obtain 3D maps of Pulsed Doppler, Color Doppler, and Power Doppler quantities in a single acquisition and revealed, at thousands of volumes per second, the complex 3D flow patterns occurring in the ventricles of the human heart during an entire cardiac cycle, as well as the 3D in vivo interaction of blood flow and wall motion during the pulse wave in the carotid at the bifurcation. This study demonstrates the potential of 3D Ultrafast Ultrasound Imaging for the 3D mapping of stiffness, tissue motion, and flow in humans in vivo and promises new clinical applications of ultrasound with reduced intra—and inter-observer variability.

  10. 3D Ultrafast Ultrasound Imaging In Vivo

    PubMed Central

    Provost, Jean; Papadacci, Clement; Arango, Juan Esteban; Imbault, Marion; Gennisson, Jean-Luc; Tanter, Mickael; Pernot, Mathieu

    2014-01-01

    Very high frame rate ultrasound imaging has recently allowed for the extension of the applications of echography to new fields of study such as the functional imaging of the brain, cardiac electrophysiology, and the quantitative real-time imaging of the intrinsic mechanical properties of tumors, to name a few, non-invasively and in real time. In this study, we present the first implementation of Ultrafast Ultrasound Imaging in three dimensions based on the use of either diverging or plane waves emanating from a sparse virtual array located behind the probe. It achieves high contrast and resolution while maintaining imaging rates of thousands of volumes per second. A customized portable ultrasound system was developed to sample 1024 independent channels and to drive a 32×32 matrix-array probe. Its capability to track in 3D transient phenomena occurring in the millisecond range within a single ultrafast acquisition was demonstrated for 3-D Shear-Wave Imaging, 3-D Ultrafast Doppler Imaging and finally 3D Ultrafast combined Tissue and Flow Doppler. The propagation of shear waves was tracked in a phantom and used to characterize its stiffness. 3-D Ultrafast Doppler was used to obtain 3-D maps of Pulsed Doppler, Color Doppler, and Power Doppler quantities in a single acquisition and revealed, for the first time, the complex 3-D flow patterns occurring in the ventricles of the human heart during an entire cardiac cycle, and the 3-D in vivo interaction of blood flow and wall motion during the pulse wave in the carotid at the bifurcation. This study demonstrates the potential of 3-D Ultrafast Ultrasound Imaging for the 3-D real-time mapping of stiffness, tissue motion, and flow in humans in vivo and promises new clinical applications of ultrasound with reduced intra- and inter-observer variability. PMID:25207828

  11. In vivo Raman spectroscopy of cervix cancers

    NASA Astrophysics Data System (ADS)

    Rubina, S.; Sathe, Priyanka; Dora, Tapas Kumar; Chopra, Supriya; Maheshwari, Amita; Krishna, C. Murali

    2014-03-01

    Cervix-cancer is the third most common female cancer worldwide. It is the leading cancer among Indian females with more than million new diagnosed cases and 50% mortality, annually. The high mortality rates can be attributed to late diagnosis. Efficacy of Raman spectroscopy in classification of normal and pathological conditions in cervix cancers on diverse populations has already been demonstrated. Our earlier ex vivo studies have shown the feasibility of classifying normal and cancer cervix tissues as well as responders/non-responders to Concurrent chemoradiotherapy (CCRT). The present study was carried out to explore feasibility of in vivo Raman spectroscopic methods in classifying normal and cancerous conditions in Indian population. A total of 182 normal and 132 tumor in vivo Raman spectra, from 63 subjects, were recorded using a fiberoptic probe coupled HE-785 spectrometer, under clinical supervision. Spectra were acquired for 5 s and averaged over 3 times at 80 mW laser power. Spectra of normal conditions suggest strong collagenous features and abundance of non-collagenous proteins and DNA in case of tumors. Preprocessed spectra were subjected to Principal Component-Linear Discrimination Analysis (PCLDA) followed by leave-one-out-cross-validation. Classification efficiency of ~96.7% and 100% for normal and cancerous conditions respectively, were observed. Findings of the study corroborates earlier studies and suggest applicability of Raman spectroscopic methods in combination with appropriate multivariate tool for objective, noninvasive and rapid diagnosis of cervical cancers in Indian population. In view of encouraging results, extensive validation studies will be undertaken to confirm the findings.

  12. In vivo light dosimetry for pleural PDT

    NASA Astrophysics Data System (ADS)

    Dimofte, Andreea; Zhu, Timothy C.; Finlay, Jarod C.; Culligan, Melissa; Edmonds, Christine E.; Friedberg, Joseph S.; Cengel, Keith; Hahn, Stephen M.

    2009-02-01

    In-vivo light Dosimetry for patients undergoing photodynamic therapy (PDT) is one of the important dosimetry quantities critical for predicting PDT outcome. This study examines the light fluence (rate) delivered to patients undergoing pleural PDT as a function of treatment time, treatment volume and surface area, and its accuracy as a function of the calibration accuracies of each isotropic detector and the calibration integrating sphere. The patients studied here were enrolled in Phase II clinical trial of Photofrin-mediated PDT for the treatment of non-small cell lung cancer with pleural effusion. The ages of the patients studied varied from 34 to 69 year old. All patients were administered 2mg per kg body weight Photoprin 24 hours before the surgery. Patients undergoing photodynamic therapy (PDT) are treated with laser light with a light fluence of 60 J/cm^2 at 630nm. Fluence rate (mW/cm^2) and cumulative fluence (J/cm^2) was monitored at 7 different sites during the entire light treatment delivery. Isotropic detectors were used for in-vivo light dosimetry. The anisotropy of each isotropic detector was found to be within 30%. The mean fluence rate delivery varied from 37.84 to 94.05 mW/cm^2 and treatment time varied from 1762 to 5232s. We have established a correlation between the treatment time and the treatment volume. The results are discussed using an integrating sphere theory and the measured tissue optical properties. The result can be used as a clinical guideline for future pleural PDT treatment.

  13. Nebivolol increases arterial distensibility in vivo.

    PubMed

    McEniery, Carmel M; Schmitt, Matthias; Qasem, Ahmad; Webb, David J; Avolio, Alberto P; Wilkinson, Ian B; Cockcroft, John R

    2004-09-01

    Arterial stiffness is a key determinant of cardiovascular risk in hypertensive patients. beta-Blockers appear to be less effective than other drugs in improving outcome in hypertensive patients, and a potential explanation may be that beta-blockers are less effective in reducing arterial stiffness. The aim of this study was to assess the direct effect of beta-blockade on pulse wave velocity (PWV), a robust measure of arterial distensibility, using a local, ovine, hind-limb model. In addition, we hypothesized that the vasodilating beta-blocker nebivolol, but not atenolol, would increase arterial distensibility in vivo. All studies were conducted in anesthetized sheep. PWV was recorded in vivo using a dual pressure-sensing catheter placed in the common iliac artery. Intraarterial infusion of nebivolol reduced PWV by 6+/-3% at the higher dose (P<0.001), but did not alter mean arterial pressure (change of -1+/-3 mm Hg, P=0.1). In contrast, atenolol had no effect on PWV (P=0.11) despite a small drop in mean pressure (change of -5+/-3 mm Hg, P<0.01). Infusion of glyceryl trinitrate led to a dose-dependent fall in PWV, and 2 nmol/min produced a similar reduction in PWV to the higher dose of nebivolol (500 nmol/min). The effect of nebivolol on PWV was significantly attenuated during coinfusion of N(G)-monomethyl-L-arginine (P=0.003) and also during coinfusion of butoxamine (P=0.02). These results demonstrate that nebivolol, but not atenolol, increases arterial distensibility. This effect of nebivolol is mediated through the release of NO via a beta2 adrenoceptor-dependent mechanism. Thus, nebivolol may be of benefit in conditions of increased large artery stiffness, such as isolated systolic hypertension. PMID:15262912

  14. Dimerization of visual pigments in vivo.

    PubMed

    Zhang, Tao; Cao, Li-Hui; Kumar, Sandeep; Enemchukwu, Nduka O; Zhang, Ning; Lambert, Alyssia; Zhao, Xuchen; Jones, Alex; Wang, Shixian; Dennis, Emily M; Fnu, Amrita; Ham, Sam; Rainier, Jon; Yau, King-Wai; Fu, Yingbin

    2016-08-01

    It is a deeply engrained notion that the visual pigment rhodopsin signals light as a monomer, even though many G protein-coupled receptors are now known to exist and function as dimers. Nonetheless, recent studies (albeit all in vitro) have suggested that rhodopsin and its chromophore-free apoprotein, R-opsin, may indeed exist as a homodimer in rod disk membranes. Given the overwhelmingly strong historical context, the crucial remaining question, therefore, is whether pigment dimerization truly exists naturally and what function this dimerization may serve. We addressed this question in vivo with a unique mouse line (S-opsin(+)Lrat(-/-)) expressing, transgenically, short-wavelength-sensitive cone opsin (S-opsin) in rods and also lacking chromophore to exploit the fact that cone opsins, but not R-opsin, require chromophore for proper folding and trafficking to the photoreceptor's outer segment. In R-opsin's absence, S-opsin in these transgenic rods without chromophore was mislocalized; in R-opsin's presence, however, S-opsin trafficked normally to the rod outer segment and produced functional S-pigment upon subsequent chromophore restoration. Introducing a competing R-opsin transmembrane helix H1 or helix H8 peptide, but not helix H4 or helix H5 peptide, into these transgenic rods caused mislocalization of R-opsin and S-opsin to the perinuclear endoplasmic reticulum. Importantly, a similar peptide-competition effect was observed even in WT rods. Our work provides convincing evidence for visual pigment dimerization in vivo under physiological conditions and for its role in pigment maturation and targeting. Our work raises new questions regarding a potential mechanistic role of dimerization in rhodopsin signaling. PMID:27462111

  15. Robert Merton Dies at 92

    ERIC Educational Resources Information Center

    Snell, Joel C.

    2006-01-01

    This article features Robert Merton, who died recently at age 92. Merton came into this world as a Jewish baby named Meyer Schkolnick. He lived in South Philly where his parents wrenched a living as blue-collar workers. Merton chose an Anglicized name to move into the Yankee dominated America of the 20's and 30's. At Harvard, he studied under…

  16. Attitudes on Death and Dying.

    ERIC Educational Resources Information Center

    Andrus, Charles E.

    This paper explored attitudes toward death and dying revealed through interviews with members of the clergy, the medical profession, funeral directors, nursing home residents, and selected others. The sampling was small and results are not intended to be representative of the groups to which these people belong. Rather, the study may be used as a…

  17. In Vivo Monitoring Program Manual, PNL-MA-574

    SciTech Connect

    Lynch, Timothy P.

    2010-07-01

    An overview of the administration for the In Vivo Monitoring Program (IVMP) for Hanford. This includes organizational structure and program responsibilities; coordination of in vivo measurements; scheduling measurements; performing measurements; reporting results; and quality assurance. Overall responsibility for the management of the IVMP rests with the Program Manager (PM). The PM is responsible for providing the required in vivo counting services for Hanford Site contractor employees in accordance with Department of Energy (DOE) requirements and the specific statements of work.

  18. Integrated Forming Simulations and Die Structural Analysis for Optimal Die Designs

    NASA Astrophysics Data System (ADS)

    Aitharaju, Venkat; Liu, Malcolm; Dong, Jennifer; Zhang, Jimmy; Wang, Chuan-tao

    2005-08-01

    After gaining a huge success in applying stamping simulations and formability analysis to validate die face developments, GM moves forward to winning total manufacturability in stamping process. Of which, ensuring die structure integrity and minimizing weight is one of the important initiatives. Stamping die design (or solid modeling of stamping dies) was traditionally conducted by following the die design manuals and standards. For any design changes beyond the standards, however, there are no math-based tools available to die designers to verify the outcome of the changes. Die structural analysis (DSA) provides a math-tool to validate the design changes and quantify the safety factors. Several years ago, GM Manufacturing Engineering — Die Center started die structural analysis to meet the increasing demands of customer needs in various areas: (1) to validate design changes; (2) to identify root cause of die breakage during the tryout and stamping operations and propose repair schemes; (3) to optimize the die design for weight reduction; (4) to improve press throughput via optimizing the scrap chute openings, and (5) to provide a math-based tool to validate revisions to the current die design standards. In the integrated forming and die structural analysis, after successful line die surface developments, the forming loads (binder force, pad force, and forming tonnages) are extracted from forming simulations and applied to solid die members for structural analyses of stress, strains, and deflections. In the past few years, Die Center conducted static, dynamic and fatigue analysis for many dies that covers the die design changes requested by die design, die construction and stamping plants. This paper presents some fundamentals and issues of integrated forming and die structural analysis and illustrates the significant impact of die structural analysis on die design, die construction and production stamping.

  19. Passive in vivo elastography from skeletal muscle noise

    SciTech Connect

    Sabra, Karim G.; Conti, Stephane; Roux, Philippe; Kuperman, W. A.

    2007-05-07

    Measuring the in vivo elastic properties of muscles (e.g., stiffness) provides a means for diagnosing and monitoring muscular activity. The authors demonstrated a passive in vivo elastography technique without an active external radiation source. This technique instead uses cross correlations of contracting skeletal muscle noise recorded with skin-mounted sensors. Each passive sensor becomes a virtual in vivo shear wave source. The results point to a low-cost, noninvasive technique for monitoring biomechanical in vivo muscle properties. The efficacy of the passive elastography technique originates from the high density of cross paths between all sensor pairs, potentially achieving the same sensitivity obtained from active elastography methods.

  20. Exploratory study on H13 steel dies

    SciTech Connect

    Sunwoo, A.J.

    1994-04-01

    Ultrahigh-strength H13 steel is a recommended die material for aluminum die casting; dies made from H13 steel can be safely water- cooled during hot working operations without cracking. However, after time the dies exhibited surface cracking and excessive wear. Erosive wear also occurs owing to high pressure injection of molten Al. An exploratory study was made of the causes for surface cracking of H13 dies. Results suggest that surface cracking is caused by interrelated factors, internal to the die material as well as externally induced conditions.

  1. Viral Vectors for in Vivo Gene Transfer

    NASA Astrophysics Data System (ADS)

    Thévenot, E.; Dufour, N.; Déglon, N.

    The transfer of DNA into the nucleus of a eukaryotic cell (gene transfer) is a central theme of modern biology. The transfer is said to be somatic when it refers to non-germline organs of a developed individual, and germline when it concerns gametes or the fertilised egg of an animal, with the aim of transmitting the relevant genetic modification to its descendents [1]. The efficient introduction of genetic material into a somatic or germline cell and the control of its expression over time have led to major advances in understanding how genes work in vivo, i.e., in living organisms (functional genomics), but also to the development of innovative therapeutic methods (gene therapy). The efficiency of gene transfer is conditioned by the vehicle used, called the vector. Desirable features for a vector are as follows: Easy to produce high titer stocks of the vector in a reproducible way. Absence of toxicity related to transduction (transfer of genetic material into the target cell, and its expression there) and no immune reaction of the organism against the vector and/or therapeutic protein. Stability in the expression of the relevant gene over time, and the possibility of regulation, e.g., to control expression of the therapeutic protein on the physiological level, or to end expression at the end of treatment. Transduction of quiescent cells should be as efficient as transduction of dividing cells. Vectors currently used fall into two categories: non-viral and viral vectors. In non-viral vectors, the DNA is complexed with polymers, lipids, or cationic detergents (described in Chap. 3). These vectors have a low risk of toxicity and immune reaction. However, they are less efficient in vivo than viral vectors when it comes to the number of cells transduced and long-term transgene expression. (Naked DNA transfer or electroporation is rather inefficient in the organism. This type of gene transfer will not be discussed here, and the interested reader is referred to the

  2. The thermal fatigue resistance of H-13 Die Steel for aluminum die casting dies

    NASA Technical Reports Server (NTRS)

    1982-01-01

    The effects of welding, five selected surface coatings, and stress relieving on the thermal fatigue resistance of H-13 Die Steel for aluminum die casting dies were studied using eleven thermal fatigue specimens. Stress relieving was conducted after each 5,000 cycle interval at 1050 F for three hours. Four thermal fatigue specimens were welded with H-13 or maraging steel welding rods at ambient and elevated temperatures and subsequently, subjected to different post-weld heat treatments. Crack patterns were examined at 5,000, 10,000, and 15,000 cycles. The thermal fatigue resistance is expressed by two crack parameters which are the average maximum crack and the average cracked area. The results indicate that a significant improvement in thermal fatigue resistance over the control was obtained from the stress-relieving treatment. Small improvements were obtained from the H-13 welded specimens and from a salt bath nitrogen and carbon-surface treatment. The other surface treatments and welded specimens either did not affect or had a detrimental influence on the thermal fatigue properties of the H-13 die steel.

  3. Stability of blocked replication forks in vivo

    PubMed Central

    Mettrick, Karla A.; Grainge, Ian

    2016-01-01

    Replication of chromosomal DNA must be carried out to completion in order for a cell to proliferate. However, replication forks can stall during this process for a variety of reasons, including nucleoprotein ‘roadblocks’ and DNA lesions. In these circumstances the replisome copying the DNA may disengage from the chromosome to allow various repair processes to restore DNA integrity and enable replication to continue. Here, we report the in vivo stability of the replication fork when it encounters a nucleoprotein blockage in Escherichia coli. Using a site-specific and reversible protein block system in conjunction with the temperature sensitive DnaC helicase loader and DnaB replicative helicase, we monitored the disappearance of the Y-shaped DNA replication fork structures using neutral-neutral 2D agarose gels. We show the replication fork collapses within 5 min of encountering the roadblock. Therefore, the stalled replication fork does not pause at a block in a stable confirmation for an extended period of time as previously postulated. PMID:26490956

  4. Translational In Vivo Models for Cardiovascular Diseases.

    PubMed

    Fliegner, Daniela; Gerdes, Christoph; Meding, Jörg; Stasch, Johannes-Peter

    2016-01-01

    Cardiovascular diseases are still the first leading cause of death and morbidity in developed countries. Experimental cardiology research and preclinical drug development in cardiology call for appropriate and especially clinically relevant in vitro and in vivo studies. The use of animal models has contributed to expand our knowledge and our understanding of the underlying mechanisms and accordingly provided new approaches focused on the improvement of diagnostic and treatment strategies of various cardiac pathologies.Numerous animal models in different species as well as in small and large animals have been developed to address cardiovascular complications, including heart failure, pulmonary hypertension, and thrombotic diseases. However, a perfect model of heart failure or other indications that reproduces every aspect of the natural disease does not exist. The complexity and heterogeneity of cardiac diseases plus the influence of genetic and environmental factors limit to mirror a particular disease with a single experimental model.Thus, drug development in the field of cardiology is not only very challenging but also inspiring; therefore animal models should be selected that reflect as best as possible the disease being investigated. Given the wide range of animal models, reflecting critical features of the human pathophysiology available nowadays increases the likelihood of the translation to the patients. Furthermore, this knowledge and the increase of the predictive value of preclinical models help us to find more efficient and reliable solutions as well as better and innovative treatment strategies for cardiovascular diseases. PMID:26552402

  5. In vivo measurement of the redox state.

    PubMed

    Praticò, D

    2001-01-01

    As part of an aerobic life, we oxidize a large pool of biomolecules to obtain chemical energy. During this process, several intermediates are formed; some are chemically unstable and are referred to as free radicals (FR). FR tend to react quickly with their surrounding biological environment; depending on the nature of the molecule attacked, different reactions can occur, i.e., lipid peroxidation, protein oxidation, or DNA oxidation products. As aerobic life has evolved, antioxidant defense systems against FR have developed. When an imbalance between production of FR (oxidants) and defense systems against them (antioxidants) happens, a situation of oxidative stress occurs. This can lead to irreversible biochemical changes, with subsequent tissue damage and disease. Establishing the involvement of FR in the pathogenesis of a disease has been difficult because of the lack of sensitive and specific methodology to detect them. No ideal biomarkers for in vivo FR-induced damage are available as yet. However, some reliable indices of FR formation are now available, and in some pathologic conditions, evidence is accumulating to show that FR damage might play a functional role. The task for the near future will be to try to simplify the analytical methodology and elucidate the molecular mechanisms underlying the formation, disposition, and kinetics of FR marker molecules. PMID:11837992

  6. Thermal Assisted In Vivo Gene Electrotransfer.

    PubMed

    Donate, Amy; Bulysheva, Anna; Edelblute, Chelsea; Jung, Derrick; Malik, Mohammad A; Guo, Siqi; Burcus, Niculina; Schoenbach, Karl; Heller, Richard

    2016-01-01

    Gene electrotransfer is an effective approach for delivering plasmid DNA to a variety of tissues. Delivery of molecules with electric pulses requires control of the electrical parameters to achieve effective delivery. Since discomfort or tissue damage may occur with high applied voltage, the reduction of the applied voltage while achieving the desired expression may be an important improvement. One possible approach is to combine electrotransfer with exogenously applied heat. Previous work performed in vitro demonstrated that increasing temperature before pulsing can enhance gene expression and made it possible to reduce electric fields while maintaining expression levels. In the study reported here, this combination was evaluated in vivo using a novel electrode device designed with an inserted laser for application of heat. The results obtained in this study demonstrated that increased temperature during electrotransfer increased expression or maintained expression with a reduction in applied voltage. With further optimization this approach may provide the basis for both a novel method and a novel instrument that may greatly enhance translation of gene electrotransfer. PMID:27029944

  7. Platelet generation in vivo and in vitro.

    PubMed

    Wang, Biao; Zheng, Jiansheng

    2016-01-01

    Platelet (PLT) transfusion, which is the primary cell therapy for thrombocytopenia, has been a source of concern in recent years due to its limitations of donor-dependent supply and soaring costs. In vitro platelet generation on an industrial scale is a possible solution requiring exploration. The technology of platelet generation ex vivo has been widely studied across the world, though the mechanisms of physiological thrombopoiesis and platelet biology function in vivo still remain elusive today. Various culture systems have been studied, most of which proved quite inefficient in generating functional platelets ex vivo, so there is still a long way to reach our ultimate goal of generating a fully functional platelet in vitro on an industrial scale. This review integrates the latest research into physiological platelet biogenesis and ex vivo-platelet/megakaryocyte (MK) generation protocols with a focus on the ability to generate PLT/MK in large quantities, summarizes current culture systems based on induced human pluripotent stem cells and adipose-derived stem cells, and discusses significant challenges that must be overcome for these approaches to be perfected. PMID:27390629

  8. Wireless Monitoring of Liver Hemodynamics In Vivo

    SciTech Connect

    Akl, Tony; Wilson, Mark A.; Ericson, Milton Nance; Farquhar, Ethan; Cote, Gerard L.

    2014-01-01

    Liver transplants have their highest technical failure rate in the first two weeks following surgery. Currently, there are limited devices for continuous, real-time monitoring of the graft. In this work, a three wavelengths system is presented that combines near-infrared spectroscopy and photoplethysmography with a processing method that can uniquely measure and separate the venous and arterial oxygen contributions. This strategy allows for the quantification of tissue oxygen consumption used to study hepatic metabolic activity and to relate it to tissue stress. The sensor is battery operated and communicates wirelessly with a data acquisition computer which provides the possibility of implantation provided sufficient miniaturization. In two in vivo porcine studies, the sensor tracked perfusion changes in hepatic tissue during vascular occlusions with a root mean square error (RMSE) of 0.135 mL/min/g of tissue. We show the possibility of using the pulsatile wave to measure the arterial oxygen saturation similar to pulse oximetry. The signal is also used to extract the venous oxygen saturation from the direct current (DC) levels. Arterial and venous oxygen saturation changes were measured with an RMSE of 2.19% and 1.39% respectively when no vascular occlusions were induced. This error increased to 2.82% and 3.83% when vascular occlusions were induced during hypoxia. These errors are similar to the resolution of a commercial oximetry catheter used as a reference. This work is the first realization of a wireless optical sensor for continuous monitoring of hepatic hemodynamics.

  9. Feasibility of in vivo cardiac HIFU ablation

    NASA Astrophysics Data System (ADS)

    Fujikura, Kana; Otsuka, Ryo; Homma, Shunichi; Muratore, Robert; Ketterling, Jeffrey A.; Lizzi, Frederic L.

    2001-05-01

    The potential for cardiac applications of HIFU remains unexamined. In order to create reproducible lesions in a beating heart, it is necessary to maintain focusing at a certain position within moving myocardial tissue. One technique is to use multiple short HIFU exposures (0.2 s) and synchronize them with an EKG signal and respiration. The left ventricular free wall (LVFW) of calf hearts were cut into 4-cm cubes, degassed in phosphate buffer saline (PBS), and heated to 37°C. An 80-mm-diam spherical-cap transducer with a focus of 90 mm was operated at a frequency of 4.67 MHz and a nominal focal point intensity of 26.9 kW/cm2. The transducer was coupled to the LVFW using degassed PBS. First, the effect of pericardial fat, focal depth, and temperature on lesion size was individually evaluated. Then the 0.2-s HIFU exposure was applied 10 to 30 times at 4-s intervals. The same HIFU transducer was applied to an open-chest canine LVFW with the same triggering protocol. Dimensions of all lesions were measured by visual examination of the fresh, unstained tissue. A histopathological examination of the lesion was also performed. The in vivo lesions were created in similar size to those in vitro.

  10. Imaging inflammatory plasma leakage in vivo.

    PubMed

    Kenne, E; Lindbom, L

    2011-05-01

    Increased vascular permeability and consequent plasma leakage from postcapillary venules is a cardinal sign of inflammation. Although the movement of plasma constituents from the vasculature to the affected tissue aids in clearing the inflammatory stimulus, excessive plasma extravasation can lead to hospitalisation or death in cases such as influenza-induced pneumonia, burns or brain injury. The use of intravital imaging has significantly contributed to the understanding of the mechanisms controlling the vascular permeability alterations that occur during inflammation. Today, intravital imaging can be performed using optical and non-optical techniques. Optical techniques, which are generally used in experimental settings, include traditional intravital fluorescence microscopy and near-infrared fluorescence imaging. Magnetic resonance (MRI) and radioisotopic imaging are used mainly in the clinical setting, but are increasingly used in experimental work, and can detect plasma leakage without optics. Although these methods are all able to visualise inflammatory plasma leakage in vivo, the spatial and temporal resolution differs between the techniques. In addition, they vary with regards to invasiveness and availability. This overview discusses the use of imaging techniques in the visualisation of inflammatory plasma leakage. PMID:21437352

  11. Optical stimulation of peripheral nerves in vivo

    NASA Astrophysics Data System (ADS)

    Wells, Jonathon D.

    This dissertation documents the emergence and validation of a new clinical tool that bridges the fields of biomedical optics and neuroscience. The research herein describes an innovative method for direct neurostimulation with pulsed infrared laser light. Safety and effectiveness of this technique are first demonstrated through functional stimulation of the rat sciatic nerve in vivo. The Holmium:YAG laser (lambda = 2.12 mum) is shown to operate at an optimal wavelength for peripheral nerve stimulation with advantages over standard electrical neural stimulation; including contact-free stimulation, high spatial selectivity, and lack of a stimulation artifact. The underlying biophysical mechanism responsible for transient optical nerve stimulation appears to be a small, absorption driven thermal gradient sustained at the axonal layer of nerve. Results explicitly prove that low frequency optical stimulation can reliably stimulate without resulting in tissue thermal damage. Based on the positive results from animal studies, these optimal laser parameters were utilized to move this research into the clinic with a combined safety and efficacy study in human subjects undergoing selective dorsal rhizotomy. The clinical Holmium:YAG laser was used to effectively stimulate human dorsal spinal roots and elicit functional muscle responses recorded during surgery without evidence of nerve damage. Overall these results predict that this technology can be a valuable clinical tool in various neurosurgical applications.

  12. Responsive corneosurfametry following in vivo skin preconditioning.

    PubMed

    Uhoda, E; Goffin, V; Pierard, G E

    2003-12-01

    Skin is subjected to many environmental threats, some of which altering the structure and function of the stratum corneum. Among them, surfactants are recognized factors that may influence irritant contact dermatitis. The present study was conducted to compare the variations in skin capacitance and corneosurfametry (CSM) reactivity before and after skin exposure to repeated subclinical injuries by 2 hand dishwashing liquids. A forearm immersion test was performed on 30 healthy volunteers. 2 daily soak sessions were performed for 5 days. At inclusion and the day following the last soak session, skin capacitance was measured and cyanoacrylate skin-surface strippings were harvested. The latter specimens were used for the ex vivo microwave CSM. Both types of assessments clearly differentiated the 2 hand dishwashing liquids. The forearm immersion test allowed the discriminant sensitivity of CSM to increase. Intact skin capacitance did not predict CSM data. By contrast, a significant correlation was found between the post-test conductance and the corresponding CSM data. In conclusion, a forearm immersion test under realistic conditions can discriminate the irritation potential between surfactant-based products by measuring skin conductance and performing CSM. In vivo skin preconditioning by surfactants increases CSM sensitivity to the same surfactants. PMID:15025702

  13. In Vivo Gait Analysis During Bone Transport.

    PubMed

    Mora-Macías, J; Reina-Romo, E; Morgaz, J; Domínguez, J

    2015-09-01

    The load bearing characteristics of the intervened limb over time in vivo are important to know in distraction osteogenesis and bone healing for the characterization of the bone maturation process. Gait analyses were performed for a group of sheep in which bone transport was carried out. The ground reaction force was measured by means of a force platform, and the gait parameters (i.e., the peak, the mean vertical ground reaction force and the impulse) were calculated during the stance phase for each limb. The results showed that these gait parameters decreased in the intervened limb and interestingly increased in the other limbs due to the implantation of the fixator. Additionally, during the process, the gait parameters exponentially approached the values for healthy animals. Corresponding radiographies showed an increasing level of ossification in the callus. This study shows, as a preliminary approach to be confirmed with more experiments, that gait analysis could be used as an alternative method to control distraction osteogenesis or bone healing. For example, these analyses could determine the appropriate time to remove the fixator. Furthermore, gait analysis has advantages over other methods because it provides quantitative data and does not require instrumented fixators. PMID:25650097

  14. Imaging axon pathfinding in zebrafish in vivo.

    PubMed

    Leung, Louis; Holt, Christine E

    2012-09-01

    Axon pathfinding in the developing animal involves a highly dynamic process in which the axonal growth cone makes continuous decisions as it navigates toward its target. Changes occurring in the growth cone with respect to retracting from or extending into complex new territories can occur in minutes. Thus, the advent of strategies to visualize axon path-finding in vivo in a live intact animal is crucial for a better understanding of how the growth cone makes such rapid decisions in response to multiple cues. Combining these strategies with loss-of-function and/or gain-of-function techniques, one can gain some insight as to which molecules are crucial to particular growth cone behaviors at specific choice points during navigation. The major advantage of using zebrafish lies in the accessibility of major axon tracts for live microscopy, as their embryonic development occurs ex utero. Furthermore, the robust embryos remain healthy during immobilization and allow for good imaging for long periods. This protocol describes the method for stabilizing and preparing live zebrafish embryos for imaging labeled axonal tracts at high spatial and temporal resolution for up to 72 h. It has been used for retinotectal axon pathfinding, but can be adapted to visualize other axon tracts of interest. PMID:22949713

  15. Imaging axon pathfinding in Xenopus in vivo.

    PubMed

    Leung, Louis; Holt, Christine E

    2012-09-01

    Axon pathfinding in the developing animal involves a highly dynamic process in which the axonal growth cone makes continuous decisions as it navigates toward its target. Changes occurring in the growth cone with respect to retracting from or extending into complex new territories can occur in minutes. Thus, the advent of strategies to visualize axon path-finding in vivo in a live intact animal is crucial for a better understanding of how the growth cone makes such rapid decisions in response to multiple cues. Combining these strategies with loss-of-function and/or gain-of-function techniques allows one to gain some insight as to which molecules are crucial to particular growth cone behaviors at specific choice points during navigation. The main advantage of using Xenopus lies in the accessibility of major axon tracts for live microscopy, as their embryonic development occurs ex utero. Furthermore, the robust embryos remain healthy during immobilization and allow for good imaging for long periods. This protocol describes the methods for stabilizing and preparing live Xenopus embryos for imaging labeled axonal tracts at high spatial and temporal resolution for up to 72 h. This approach can been used to investigate how the knockdown of certain gene functions can affect the speed of navigation through the well-studied Xenopus retinotectal pathway. It can be adapted to visualize other axon tracts of interest. PMID:22949712

  16. Difficulty in dislodging in vivo fixed radiostrontium.

    PubMed

    Sonawane, V R; Jagtap, V S; Pahuja, D N; Rajan, M G R; Samuel, A M

    2004-07-01

    Many trials based on the basic phenomena of isotopic dilution, adsorption, ion exchange, chelation, etc., have been attempted for the decorporation of radiostrontium, particularly Sr, after its entry in the in vivo system. We have recently demonstrated a non-isotopic carrier effect of some common calcium salts (calcium = 9 mg mL) to reduce the whole body retention of radiostrontium, if administered within 2 h after radiostrontium exposure and furthermore once daily, in rats, supplemented with calcium fortified diet. However, 25-30% of radiostrontium (compared to 50-60% in untreated animals) was still found to be retained in the animal even after 2 wk of treatment. Trial of some simple interventional measures, which would not adversely affect the animal metabolism, like pyrophosphate and magnesium sulfate, sodium citrate, chitin (a bio-absorbent), crown ether (a metal-chelator), and ammonium chloride, was therefore attempted to dislodge this remaining radiostrontium by switching over these animals to normal diet and subjecting them to different lines of treatment with these simple interventions through diet and drinking water separately for a further 4 wk. However, this remaining portion of radiostrontium is fixed in the bone and is difficult to dislodge. PMID:15194921

  17. Macrophage plasticity and polarization: in vivo veritas

    PubMed Central

    Sica, Antonio; Mantovani, Alberto

    2012-01-01

    Diversity and plasticity are hallmarks of cells of the monocyte-macrophage lineage. In response to IFNs, Toll-like receptor engagement, or IL-4/IL-13 signaling, macrophages undergo M1 (classical) or M2 (alternative) activation, which represent extremes of a continuum in a universe of activation states. Progress has now been made in defining the signaling pathways, transcriptional networks, and epigenetic mechanisms underlying M1-M2 or M2-like polarized activation. Functional skewing of mononuclear phagocytes occurs in vivo under physiological conditions (e.g., ontogenesis and pregnancy) and in pathology (allergic and chronic inflammation, tissue repair, infection, and cancer). However, in selected preclinical and clinical conditions, coexistence of cells in different activation states and unique or mixed phenotypes have been observed, a reflection of dynamic changes and complex tissue-derived signals. The identification of mechanisms and molecules associated with macrophage plasticity and polarized activation provides a basis for macrophage-centered diagnostic and therapeutic strategies. PMID:22378047

  18. In vivo photoacoustic imaging of mouse embryos

    NASA Astrophysics Data System (ADS)

    Laufer, Jan; Norris, Francesca; Cleary, Jon; Zhang, Edward; Treeby, Bradley; Cox, Ben; Johnson, Peter; Scambler, Pete; Lythgoe, Mark; Beard, Paul

    2012-06-01

    The ability to noninvasively image embryonic vascular anatomy in mouse models is an important requirement for characterizing the development of the normal cardiovascular system and malformations in the heart and vascular supply. Photoacoustic imaging, which can provide high resolution non invasive images of the vasculature based upon optical absorption by endogenous hemoglobin, is well suited to this application. In this study, photoacoustic images of mouse embryos were obtained ex vivo and in vivo. The images show intricate details of the embryonic vascular system to depths of up to 10 mm, which allowed whole embryos to be imaged in situ. To achieve this, an all-optical photoacoustic scanner and a novel time reversal image reconstruction algorithm, which provide deep tissue imaging capability while maintaining high spatial resolution and contrast were employed. This technology may find application as an imaging tool for preclinical embryo studies in developmental biology as well as more generally in preclinical and clinical medicine for studying pathologies characterized by changes in the vasculature.

  19. Astrocytes regulate cortical state switching in vivo

    PubMed Central

    Poskanzer, Kira E.; Yuste, Rafael

    2016-01-01

    The role of astrocytes in neuronal function has received increasing recognition, but disagreement remains about their function at the circuit level. Here we use in vivo two-photon calcium imaging of neocortical astrocytes while monitoring the activity state of the local neuronal circuit electrophysiologically and optically. We find that astrocytic calcium activity precedes spontaneous circuit shifts to the slow-oscillation–dominated state, a neocortical rhythm characterized by synchronized neuronal firing and important for sleep and memory. Further, we show that optogenetic activation of astrocytes switches the local neuronal circuit to this slow-oscillation state. Finally, using two-photon imaging of extracellular glutamate, we find that astrocytic transients in glutamate co-occur with shifts to the synchronized state and that optogenetically activated astrocytes can generate these glutamate transients. We conclude that astrocytes can indeed trigger the low-frequency state of a cortical circuit by altering extracellular glutamate, and therefore play a causal role in the control of cortical synchronizations. PMID:27122314

  20. Controlled cobalt doping of magnetosomes in vivo.

    PubMed

    Staniland, Sarah; Williams, Wyn; Telling, Neil; Van Der Laan, Gerrit; Harrison, Andrew; Ward, Bruce

    2008-03-01

    Magnetotactic bacteria biomineralize iron into magnetite (Fe3O4) nanoparticles that are surrounded by lipid vesicles. These 'magnetosomes' have considerable potential for use in bio- and nanotechnological applications because of their narrow size and shape distribution and inherent biocompatibility. The ability to tailor the magnetic properties of magnetosomes by chemical doping would greatly expand these applications; however, the controlled doping of magnetosomes has so far not been achieved. Here, we report controlled in vivo cobalt doping of magnetosomes in three strains of the bacterium Magnetospirillum. The presence of cobalt increases the coercive field of the magnetosomes--that is, the field necessary to reverse their magnetization--by 36-45%, depending on the strain and the cobalt content. With elemental analysis, X-ray absorption and magnetic circular dichroism, we estimate the cobalt content to be between 0.2 and 1.4%. These findings provide an important advance in designing biologically synthesized nanoparticles with useful highly tuned magnetic properties. PMID:18654488

  1. Astrocytes regulate cortical state switching in vivo.

    PubMed

    Poskanzer, Kira E; Yuste, Rafael

    2016-05-10

    The role of astrocytes in neuronal function has received increasing recognition, but disagreement remains about their function at the circuit level. Here we use in vivo two-photon calcium imaging of neocortical astrocytes while monitoring the activity state of the local neuronal circuit electrophysiologically and optically. We find that astrocytic calcium activity precedes spontaneous circuit shifts to the slow-oscillation-dominated state, a neocortical rhythm characterized by synchronized neuronal firing and important for sleep and memory. Further, we show that optogenetic activation of astrocytes switches the local neuronal circuit to this slow-oscillation state. Finally, using two-photon imaging of extracellular glutamate, we find that astrocytic transients in glutamate co-occur with shifts to the synchronized state and that optogenetically activated astrocytes can generate these glutamate transients. We conclude that astrocytes can indeed trigger the low-frequency state of a cortical circuit by altering extracellular glutamate, and therefore play a causal role in the control of cortical synchronizations. PMID:27122314

  2. Guide for extrusion dies eliminates straightening operation

    NASA Technical Reports Server (NTRS)

    Gyorgak, C. A.; Hoover, R. J.

    1964-01-01

    To prevent distortion of extruded metal, a guidance assembly is aligned with the die. As the metal emerges from the extrusion dies, it passes directly into the receiver and straightening tube system, and the completed extrusion is withdrawn.

  3. [Dying with cancer: Hollywood lessons].

    PubMed

    Niemeyer, Fernanda; Kruse, Maria Henriqueta Luce

    2013-12-01

    The study attempts to understand how dying from cancer is portrayed by five movies produced in Hollywood between 1993 and 2006. Based on the cultural studies and their post-structuralism version and supported by the notions of discourse and subjectivity, as proposed by philosopher Michel Foucault, we suggest one of the possible readings of the movie picture corpus. We assess how the movie picture discourse acts as a cultural pedagogy that produces ways of seeing dying with cancer: immortalizing the healthy body image, silencing death, taking care of the dead body and, finally, accepting death. Our proposal is intended to stimulate reflections that may contribute to care and education in nursing. PMID:25080714

  4. In vivo and in vitro mixture modeling of endocrine disruptors

    EPA Science Inventory

    Humans, fish and wildlife are exposed to more than one chemical at a time. There is concern over the potential effects of exposure to mixtures of EDs. We have conducted invitro and in vivo studies to determine how EDs in mixtures interact. Our in vivo studies have examined the ef...

  5. In Vivo Target Validation Using Biological Molecules in Drug Development.

    PubMed

    Sim, Derek S; Kauser, Katalin

    2016-01-01

    Drug development is a resource-intensive process requiring significant financial and time investment. Preclinical target validation studies and in vivo testing of the therapeutic molecules in clinically relevant disease models can accelerate and significantly de-risk later stage clinical development. In this chapter, we will focus on (1) in vivo animal models and (2) pharmacological tools for target validation. PMID:26552401

  6. Killing, letting die and euthanasia.

    PubMed

    Husak, D N

    1979-12-01

    Medical ethicists debate whether or not the moral assessment of cases of euthanasia should depend on whether the patient is 'killed' or 'allowed to die'. The usual presupposition is that a clear distinction between killing and letting die can be drawn so that this substantive question is not begged. I contend that the categorisation of cases of instances of killing rather than as instances of letting die depends in part on a prior moral assessment of the case. Hence is it trivially rather than substantively true that the distinction has moral significance. But even if a morally neutral (ie non-question begging) distinction could be drawn, its application to the euthanasia controversy is problematic. I illustrate the difficulties of employing this distinction to reach moral conclusions by critically discussing Philippa Foot's recent treatment of euthanasia. I conclude that even if an act of euthanasia is an instance of killing, and there exists a prima facie moral duty not to kill, and no more stringent duty overrides this duty, one still cannot determine such an act to be morally impermissible. PMID:541821

  7. Adolescents’ Perceived Risk of Dying

    PubMed Central

    Fischhoff, Baruch; de Bruin, Wändi Bruine; Parker, Andrew M.; Millstein, Susan G.; Halpern-Felsher, Bonnie L.

    2009-01-01

    Purpose Although adolescents’ expectations are accurate or moderately optimistic for many significant life events, they greatly overestimate their chances of dying soon. We examine here whether adolescents’ mortality judgments are correlated with their perceptions of direct threats to their survival. Such sensitivity would indicate the importance of ensuring that adolescents have accurate information about those threats, as well as the psychological support needed to deal with them. Methods Data from two separate studies were used: a national study of 3,436 14–18 year old adolescents and a regional sample of 124 7th graders and 132 9th graders, 12–16 years old. Participants were asked about their chance of dying in the next year and before age 20, and about the extent of various threats to their physical well being. Results Adolescents in both samples greatly overestimated their chance of dying. Those mortality estimates were higher for adolescents who reported direct threats (e.g., an unsafe neighborhood). Thus, adolescents were sensitive to the relative size of threats to their survival, but not to the implications for absolute risk levels. Conclusions Contrary to the folk wisdom that adolescents have a unique sense of invulnerability, those studied here reported an exaggerated sense of mortality, which was highest among those reporting greater threats in their lives. Such fears could affect adolescents’ short-term well being and future planning. PMID:20159504

  8. Contoured Orifice for Silicon-Ribbon Die

    NASA Technical Reports Server (NTRS)

    Mackintosh, B. H.

    1985-01-01

    Die configuration encourages purity and stable growth. Contour of die orifice changes near ribbon edges. As result, silicon ribbon has nearly constant width and little carbon contamination. Die part of furnace being developed to produce high-quality, low-cost material for solar cells.

  9. Jewish tradition in death and dying.

    PubMed

    Ross, H M

    1998-10-01

    Death is often a spiritually difficult time for the dying and their families. Judaism approaches dying with some unique views that can differ from other religious traditions. Through an understanding of Jewish tradition, nurses can ease the dying process for Jewish patients and their families. PMID:10036429

  10. Cross-linking Measurements of In Vivo Protein Complex Topologies*

    PubMed Central

    Zheng, Chunxiang; Yang, Li; Hoopmann, Michael R.; Eng, Jimmy K.; Tang, Xiaoting; Weisbrod, Chad R.; Bruce, James E.

    2011-01-01

    Identification and measurement of in vivo protein interactions pose critical challenges in the goal to understand biological systems. The measurement of structures and topologies of proteins and protein complexes as they exist in cells is particularly challenging, yet critically important to improve understanding of biological function because proteins exert their intended function only through the structures and interactions they exhibit in vivo. In the present study, protein interactions in E. coli cells were identified in our unbiased cross-linking approach, yielding the first in vivo topological data on many interactions and the largest set of identified in vivo cross-linked peptides produced to date. These data show excellent agreement with protein and complex crystal structures where available. Furthermore, our unbiased data provide novel in vivo topological information that can impact understanding of biological function, even for cases where high resolution structures are not yet available. PMID:21697552

  11. Multifunctional in vivo imaging of pancreatic islets during diabetes development.

    PubMed

    Li, Ge; Wu, Binlin; Ward, Meliza G; Chong, Angie C N; Mukherjee, Sushmita; Chen, Shuibing; Hao, Mingming

    2016-07-15

    Pancreatic islet dysfunction leading to insufficient glucose-stimulated insulin secretion triggers the clinical onset of diabetes. How islet dysfunction develops is not well understood at the cellular level, partly owing to the lack of approaches to study single islets longitudinally in vivo Here, we present a noninvasive, high-resolution system to quantitatively image real-time glucose metabolism from single islets in vivo, currently not available with any other method. In addition, this multifunctional system simultaneously reports islet function, proliferation, vasculature and macrophage infiltration in vivo from the same set of images. Applying our method to a longitudinal high-fat diet study revealed changes in islet function as well as alternations in islet microenvironment. More importantly, this label-free system enabled us to image real-time glucose metabolism directly from single human islets in vivo for the first time, opening the door to noninvasive longitudinal in vivo studies of healthy and diabetic human islets. PMID:27270669

  12. Noncanonical autophagy inhibits the autoinflammatory, lupus-like response to dying cells.

    PubMed

    Martinez, Jennifer; Cunha, Larissa D; Park, Sunmin; Yang, Mao; Lu, Qun; Orchard, Robert; Li, Quan-Zhen; Yan, Mei; Janke, Laura; Guy, Cliff; Linkermann, Andreas; Virgin, Herbert W; Green, Douglas R

    2016-05-01

    Defects in clearance of dying cells have been proposed to underlie the pathogenesis of systemic lupus erythematosus (SLE). Mice lacking molecules associated with dying cell clearance develop SLE-like disease, and phagocytes from patients with SLE often display defective clearance and increased inflammatory cytokine production when exposed to dying cells in vitro. Previously, we and others described a form of noncanonical autophagy known as LC3-associated phagocytosis (LAP), in which phagosomes containing engulfed particles, including dying cells, recruit elements of the autophagy pathway to facilitate maturation of phagosomes and digestion of their contents. Genome-wide association studies have identified polymorphisms in the Atg5 (ref. 8) and possibly Atg7 (ref. 9) genes, involved in both canonical autophagy and LAP, as markers of a predisposition for SLE. Here we describe the consequences of defective LAP in vivo. Mice lacking any of several components of the LAP pathway show increased serum levels of inflammatory cytokines and autoantibodies, glomerular immune complex deposition, and evidence of kidney damage. When dying cells are injected into LAP-deficient mice, they are engulfed but not efficiently degraded and trigger acute elevation of pro-inflammatory cytokines but not anti-inflammatory interleukin (IL)-10. Repeated injection of dying cells into LAP-deficient, but not LAP-sufficient, mice accelerated the development of SLE-like disease, including increased serum levels of autoantibodies. By contrast, mice deficient in genes required for canonical autophagy but not LAP do not display defective dying cell clearance, inflammatory cytokine production, or SLE-like disease, and, like wild-type mice, produce IL-10 in response to dying cells. Therefore, defects in LAP, rather than canonical autophagy, can cause SLE-like phenomena, and may contribute to the pathogenesis of SLE. PMID:27096368

  13. Characterization of Bacillus anthracis Persistence In Vivo

    PubMed Central

    Jenkins, Sarah A.; Xu, Yi

    2013-01-01

    Pulmonary exposure to Bacillus anthracis spores initiates inhalational anthrax, a life-threatening infection. It is known that dormant spores can be recovered from the lungs of infected animals months after the initial spore exposure. Consequently, a 60-day course antibiotic treatment is recommended for exposed individuals. However, there has been little information regarding details or mechanisms of spore persistence in vivo. In this study, we investigated spore persistence in a mouse model. The results indicated that weeks after intranasal inoculation with B. anthracis spores, substantial amounts of spores could be recovered from the mouse lung. Moreover, spores of B. anthracis were significantly better at persisting in the lung than spores of a non-pathogenic Bacillus subtilis strain. The majority of B. anthracis spores in the lung were tightly associated with the lung tissue, as they could not be readily removed by lavage. Immunofluorescence staining of lung sections showed that spores associated with the alveolar and airway epithelium. Confocal analysis indicated that some of the spores were inside epithelial cells. This was further confirmed by differential immunofluorescence staining of lung cells harvested from the infected lungs, suggesting that association with lung epithelial cells may provide an advantage to spore persistence in the lung. There was no or very mild inflammation in the infected lungs. Furthermore, spores were present in the lung tissue as single spores rather than in clusters. We also showed that the anthrax toxins did not play a role in persistence. Together, the results suggest that B. anthracis spores have special properties that promote their persistence in the lung, and that there may be multiple mechanisms contributing to spore persistence. PMID:23750280

  14. Wireless monitoring of liver hemodynamics in vivo.

    PubMed

    Akl, Tony J; Wilson, Mark A; Ericson, M Nance; Farquhar, Ethan; Coté, Gerard L

    2014-01-01

    Liver transplants have their highest technical failure rate in the first two weeks following surgery. Currently, there are limited devices for continuous, real-time monitoring of the graft. In this work, a three wavelengths system is presented that combines near-infrared spectroscopy and photoplethysmography with a processing method that can uniquely measure and separate the venous and arterial oxygen contributions. This strategy allows for the quantification of tissue oxygen consumption used to study hepatic metabolic activity and to relate it to tissue stress. The sensor is battery operated and communicates wirelessly with a data acquisition computer which provides the possibility of implantation provided sufficient miniaturization. In two in vivo porcine studies, the sensor tracked perfusion changes in hepatic tissue during vascular occlusions with a root mean square error (RMSE) of 0.135 mL/min/g of tissue. We show the possibility of using the pulsatile wave to measure the arterial oxygen saturation similar to pulse oximetry. The signal is also used to extract the venous oxygen saturation from the direct current (DC) levels. Arterial and venous oxygen saturation changes were measured with an RMSE of 2.19% and 1.39% respectively when no vascular occlusions were induced. This error increased to 2.82% and 3.83% when vascular occlusions were induced during hypoxia. These errors are similar to the resolution of a commercial oximetry catheter used as a reference. This work is the first realization of a wireless optical sensor for continuous monitoring of hepatic hemodynamics. PMID:25019160

  15. Wireless Monitoring of Liver Hemodynamics In Vivo

    PubMed Central

    Akl, Tony J.; Wilson, Mark A.; Ericson, M. Nance; Farquhar, Ethan; Coté, Gerard L.

    2014-01-01

    Liver transplants have their highest technical failure rate in the first two weeks following surgery. Currently, there are limited devices for continuous, real-time monitoring of the graft. In this work, a three wavelengths system is presented that combines near-infrared spectroscopy and photoplethysmography with a processing method that can uniquely measure and separate the venous and arterial oxygen contributions. This strategy allows for the quantification of tissue oxygen consumption used to study hepatic metabolic activity and to relate it to tissue stress. The sensor is battery operated and communicates wirelessly with a data acquisition computer which provides the possibility of implantation provided sufficient miniaturization. In two in vivo porcine studies, the sensor tracked perfusion changes in hepatic tissue during vascular occlusions with a root mean square error (RMSE) of 0.135 mL/min/g of tissue. We show the possibility of using the pulsatile wave to measure the arterial oxygen saturation similar to pulse oximetry. The signal is also used to extract the venous oxygen saturation from the direct current (DC) levels. Arterial and venous oxygen saturation changes were measured with an RMSE of 2.19% and 1.39% respectively when no vascular occlusions were induced. This error increased to 2.82% and 3.83% when vascular occlusions were induced during hypoxia. These errors are similar to the resolution of a commercial oximetry catheter used as a reference. This work is the first realization of a wireless optical sensor for continuous monitoring of hepatic hemodynamics. PMID:25019160

  16. In Vivo Imaging of Human Neuroinflammation.

    PubMed

    Albrecht, Daniel S; Granziera, Cristina; Hooker, Jacob M; Loggia, Marco L

    2016-04-20

    Neuroinflammation is implicated in the pathophysiology of a growing number of human disorders, including multiple sclerosis, chronic pain, traumatic brain injury, and amyotrophic lateral sclerosis. As a result, interest in the development of novel methods to investigate neuroinflammatory processes, for the purpose of diagnosis, development of new therapies, and treatment monitoring, has surged over the past 15 years. Neuroimaging offers a wide array of non- or minimally invasive techniques to characterize neuroinflammatory processes. The intent of this Review is to provide brief descriptions of currently available neuroimaging methods to image neuroinflammation in the human central nervous system (CNS) in vivo. Specifically, because of the relatively widespread accessibility of equipment for nuclear imaging (positron emission tomography [PET]; single photon emission computed tomography [SPECT]) and magnetic resonance imaging (MRI), we will focus on strategies utilizing these technologies. We first provide a working definition of "neuroinflammation" and then discuss available neuroimaging methods to study human neuroinflammatory processes. Specifically, we will focus on neuroimaging methods that target (1) the activation of CNS immunocompetent cells (e.g. imaging of glial activation with TSPO tracer [(11)C]PBR28), (2) compromised BBB (e.g. identification of MS lesions with gadolinium-enhanced MRI), (3) CNS-infiltration of circulating immune cells (e.g. tracking monocyte infiltration into brain parenchyma with iron oxide nanoparticles and MRI), and (4) pathological consequences of neuroinflammation (e.g. imaging apoptosis with [(99m)Tc]Annexin V or iron accumulation with T2* relaxometry). This Review provides an overview of state-of-the-art techniques for imaging human neuroinflammation which have potential to impact patient care in the foreseeable future. PMID:26985861

  17. Optimized In Vivo Transfer of Small Interfering RNA Targeting Dermal Tissue Using In Vivo Surface Electroporation

    PubMed Central

    Broderick, Kate E; Chan, Amy; Lin, Feng; Shen, Xuefei; Kichaev, Gleb; Khan, Amir S; Aubin, Justin; Zimmermann, Tracy S; Sardesai, Niranjan Y.

    2012-01-01

    Electroporation (EP) of mammalian tissue is a technique that has been used successfully in the clinic for the delivery of genetic-based vaccines in the form of DNA plasmids. There is great interest in platforms which efficiently deliver RNA molecules such as messenger RNA and small interfering RNA (siRNA) to mammalian tissue. However, the in vivo delivery of RNA enhanced by EP has not been extensively characterized. This paper details the optimization of electrical parameters for a novel low-voltage EP method to deliver oligonucleotides (both DNA and RNA) to dermal tissue in vivo. Initially, the electrical parameters were optimized for dermal delivery of plasmid DNA encoding green fluorescent protein (GFP) using this novel surface dermal EP device. While all investigated parameters resulted in visible transfection, voltage parameters in the 10 V range elicited the most robust signal. The parameters optimized for DNA, were then assessed for translation of successful electrotransfer of siRNA into dermal tissue. Robust tagged-siRNA transfection in skin was detected. We then assessed whether these parameters translated to successful transfer of siRNA resulting in gene knockdown in vivo. Using a reporter gene construct encoding GFP and tagged siRNA targeting the GFP message, we show simultaneous transfection of the siRNA to the skin via EP and the concomitant knockdown of the reporter gene signal. The siRNA delivery was accomplished with no evidence of injection site inflammation or local tissue damage. The minimally invasive low-voltage EP method is thus capable of efficiently delivering both DNA and RNA molecules to dermal tissue in a tolerable manner. PMID:23344722

  18. Optimized in vivo transfer of small interfering RNA targeting dermal tissue using in vivo surface electroporation.

    PubMed

    Broderick, Kate E; Chan, Amy; Lin, Feng; Shen, Xuefei; Kichaev, Gleb; Khan, Amir S; Aubin, Justin; Zimmermann, Tracy S; Sardesai, Niranjan Y

    2012-01-01

    Electroporation (EP) of mammalian tissue is a technique that has been used successfully in the clinic for the delivery of genetic-based vaccines in the form of DNA plasmids. There is great interest in platforms which efficiently deliver RNA molecules such as messenger RNA and small interfering RNA (siRNA) to mammalian tissue. However, the in vivo delivery of RNA enhanced by EP has not been extensively characterized. This paper details the optimization of electrical parameters for a novel low-voltage EP method to deliver oligonucleotides (both DNA and RNA) to dermal tissue in vivo. Initially, the electrical parameters were optimized for dermal delivery of plasmid DNA encoding green fluorescent protein (GFP) using this novel surface dermal EP device. While all investigated parameters resulted in visible transfection, voltage parameters in the 10 V range elicited the most robust signal. The parameters optimized for DNA, were then assessed for translation of successful electrotransfer of siRNA into dermal tissue. Robust tagged-siRNA transfection in skin was detected. We then assessed whether these parameters translated to successful transfer of siRNA resulting in gene knockdown in vivo. Using a reporter gene construct encoding GFP and tagged siRNA targeting the GFP message, we show simultaneous transfection of the siRNA to the skin via EP and the concomitant knockdown of the reporter gene signal. The siRNA delivery was accomplished with no evidence of injection site inflammation or local tissue damage. The minimally invasive low-voltage EP method is thus capable of efficiently delivering both DNA and RNA molecules to dermal tissue in a tolerable manner. PMID:23344722

  19. Percutaneous Valve Replacement: Significance of Different Delivery Systems In Vitro and In Vivo

    SciTech Connect

    Attmann, Tim; Lutter, Georg Quaden, Rene; Jahnke, Thomas; Rumberg, Kristin; Cremer, Jochen; Muller-Hulsbeck, Stefan

    2006-06-15

    Background and purpose. Percutaneous heart valve replacement is an exciting growing field in cardiovascular medicine yet still with some major problems. Only sophisticated improvement of the instruments could make it a real alternative to conventional surgery. Therefore, the aim of this study was to evaluate different delivery devices for percutaneous heart valve replacement in vitro and in vivo. Methods. A catheter prototype designed by our group, and two commercially available devices for the delivery of esophageal stents and aortic endoprostheses, were tested. After in vitro experiments, an ovine animal model of transfemoral pulmonary valve implantation was established using biological valved self-expanding stents. Only the delivery device for aortic endografts (Medtronic, Talent, Santa Rosa, CA, USA) allowed fast in vitro procedures without material fatigue. This device was chosen for the in vivo tests. Results. Technical success was achieved in 9 of 10 animals (90%). One animal died after perforation of the ventricular wall. Orthotopic pulmonary placement was performed in 6 animals and intentional supravalvular valved stent placement in 3 animals. Conclusions. An adequate in vitro model for this evolving field of interventional heart valve replacement is presented. Furthermore, the present study pinpoints the key characteristics that are mandatory for a delivery system in percutaneous pulmonary valve implantation. With regard to the delivery device's ductility observed during this 'venous' study, an approach to transfemoral aortic valve implantation seems feasible.

  20. Viral Nanoparticles for In vivo Tumor Imaging

    PubMed Central

    Wen, Amy M.; Lee, Karin L.; Yildiz, Ibrahim; Bruckman, Michael A.; Shukla, Sourabh; Steinmetz, Nicole F.

    2012-01-01

    proteinaceous nanoparticles while enhancing their pharmacokinetics 8,11. We demonstrate tumor homing of PEGylated VNPs using a mouse xenograft tumor model. A combination of fluorescence imaging of tissues ex vivo using Maestro Imaging System, fluorescence quantification in homogenized tissues, and confocal microscopy is used to study biodistribution. VNPs are cleared via the reticuloendothelial system (RES); tumor homing is achieved passively via the enhanced permeability and retention (EPR) effect12. The VNP nanotechnology is a powerful plug-and-play technology to image and treat sites of disease in vivo. We are further developing VNPs to carry drug cargos and clinically-relevant imaging moieties, as well as tissue-specific ligands to target molecular receptors overexpressed in cancer and cardiovascular disease. PMID:23183850

  1. Die Welt des Herrn Kuhn

    NASA Astrophysics Data System (ADS)

    Kern, Daniela

    Eines Morgens erwachte Herr Kuhn fröstelnd und staunte darüber, dass es in seinerWohnung eiskalt war. Dennoch quälte er sich aus seiner kuscheligen Bettdecke heraus und schlurfte ins Bad. "Hoffentlich wird wenigstens das Wasser warm", dachte er sich, als er den Wasserhahn betätigte - aber es kam nicht nur kein warmesWasser, außer einem unheilvollen Gluckser kam gar nichts aus der Leitung. "Dann werde ich wohl mal den Klempner anrufen", sprach er sich leise in den Bart und griff zu seinem Handy - doch das Netz war tot! Herr Kuhn begann nun, sich ernsthaft Sorgen zu machen, "Oje, was ist denn heute nur los? Ist irgendetwas Schlimmes passiert?" Um einen besseren Überblick über die Lage zu bekommen und sich austauschen zu können, brannte er nun förmlich darauf, rauszugehen und zur Arbeit zu fahren. An anderen Tagen, die er frisch geduscht und mit Kaffee und Marmeladen-Brot begann, war er selten so motiviert. So ging er also nun mit leerem Magen aus dem Haus. Hätte er den Versuch unternommen, sein tägliches Marmeladenbrot zuzubereiten, und dafür den Kühlschrank geöffnet, um das Marmeladenglas herauszunehmen, wäre ihm aufgefallen, dass auch die Stromversorgung Störungen unterworfen war, unschön zu erkennen an den ersten grünen, felligen Inseln auf seinem Lieblingskäse.

  2. Contamination of in vivo bone-lead measurements

    NASA Astrophysics Data System (ADS)

    Todd, A. C.

    2000-01-01

    This paper addresses one aspect of the calibration of a 109 Cd-based K-shell x-ray fluorescence measurement system, namely the treatment of the calibration line intercept. Under circumstances of contamination, the intercept may warrant statistical treatment different from that currently performed. This paper proposes refinements to the existing method of subtracting the phantom calibration line intercept from in vivo responses in the calculation of in vivo concentrations. These refinements are recommended because the existing method can underestimate in vivo concentrations by a small amount under normal operating conditions. Contamination of the calibration standard matrix of plaster of Paris by both lead and non-lead contaminants is addressed.

  3. Plasma and Cavitation Dynamics during Pulsed Laser Microsurgery in vivo

    SciTech Connect

    Hutson, M. Shane; Ma Xiaoyan

    2007-10-12

    We compare the plasma and cavitation dynamics underlying pulsed laser microsurgery in water and in fruit fly embryos (in vivo)--specifically for nanosecond pulses at 355 and 532 nm. We find two key differences. First, the plasma-formation thresholds are lower in vivo --especially at 355 nm--due to the presence of endogenous chromophores that serve as additional sources for plasma seed electrons. Second, the biological matrix constrains the growth of laser-induced cavitation bubbles. Both effects reduce the disrupted region in vivo when compared to extrapolations from measurements in water.

  4. In Vivo Metal Ion Imaging Using Fluorescent Sensors.

    PubMed

    Van de Bittner, Genevieve C; Hirayama, Tasuku

    2016-01-01

    In vivo imaging in living animals provides the ability to monitor alterations of signaling molecules, ions, and other biological components during various life stages and in disease. The data gained from in vivo imaging can be used for biological discovery or to determine elements of disease progression and can inform the development and translation of therapeutics. Herein, we present theories behind small-molecule, fluorescent, metal ion sensors as well as the methods for their successful application to in vivo metal ion imaging, including ex vivo validation. PMID:27283424

  5. Label-free optical activation of astrocyte in vivo

    NASA Astrophysics Data System (ADS)

    Choi, Myunghwan; Yoon, Jonghee; Ku, Taeyun; Choi, Kyungsun; Choi, Chulhee

    2011-07-01

    As the most abundant cell type in the central nervous system, astrocyte has been one of main research topics in neuroscience. Although various tools have been developed, at present, there is no tool that allows noninvasive activation of astrocyte in vivo without genetic or pharmacological perturbation. Here we report a noninvasive label-free optical method for physiological astrocyte activation in vivo using a femtosecond pulsed laser. We showed the laser stimulation robustly induced astrocytic calcium activation in vivo and further verified physiological relevance of the calcium increase by demonstrating astrocyte mediated vasodilation in the brain. This novel optical method will facilitate noninvasive physiological study on astrocyte function.

  6. Vascular cell biology in vivo: a new piscine paradigm?

    PubMed

    Weinstein, Brant

    2002-09-01

    Understanding how blood vessels form has become increasingly important in recent years yet remains difficult to study. The architecture and context of blood vessels are difficult to reproduce in vitro, and most developing blood vessels in vivo are relatively inaccessible to observation and experimental manipulation. Zebrafish, however, provide several advantages. They have small, accessible, transparent embryos and larvae, facilitating high-resolution imaging in vivo. In addition, genetic and experimental tools and methods are available for functional manipulation of the entire organism, vascular tissues or even single vascular- or non-vascular cells. Together, these features make the fish amenable to 'in vivo vascular cell biology'. PMID:12220865

  7. Sequestrated Thrombolysis: Comparative Evaluation In Vivo

    SciTech Connect

    Roy, Sumit; Laerum, Frode; Brosstad, Frank; Kvernebo, Knut; Sakariassen, Kjell S.

    2000-03-15

    Purpose: Lysis of a thrombus is a function of the local concentration of thrombolytic enzymes. This study was designed to determine in a porcine model of acute deep vein thrombosis (DVT) whether perithrombic sequestration of small volumes of a concentrated enzyme solution can accelerate the process of thrombolysis.Methods: DVT was induced in both hind limbs using a previously described technique (n = 32). Thirty minutes later the animal was heparinized and unilateral thrombolysis was attempted using 8 mg recombinant tissue plasminogen activator (rt-PA); saline was administered in the opposite leg. For conventional high-volume infusion (CI) (n = 5) rt-PA (0.067 mg/ml) was infused at 1 ml/min. For sequestrated thrombolysis the external iliac vein was endoluminally occluded, and rt-PA (0.25 mg/ml) administered either for proximal injection (ST-P) (n = 5), as a bolus every 3 min through a microcatheter placed via the balloon catheter, or for transthrombic injection (ST-T) (n = 5), as a bolus every 3 min through a Katzen wire in the balloon catheter. At autopsy, the thrombus mass in the iliofemoral veins was measured, and the extent of residual thrombosis in the venous tributaries graded at four sites. From these data a thrombolysis score was calculated.Results: One pig died before thrombolysis could be performed. Only with ST-T was residual thrombus mass in the test limb normalized to control, residual thrombus index (RTI), consistently less than unity. The median RTI of this group was 0.50 (range 0.39-0.97) compared with 1.22 (0.64-1.38) for ST-P and 0.88 (0.37-1.13) for CI. Compared with contralateral controls, a lower grade of residual thrombosis in tributaries was observed in test limbs at more venous sites with ST-T (8/20; 95% confidence interval 5-13) and ST-P (9/20; confidence interval 5-13) than with CI (2/20; confidence interval 0-5) (p= 0.04). A trend toward lower thrombolysis scores was observed with ST-T (p = 0.08). Systemic fibrinogenolysis was not

  8. Evaluation of permanent die coatings to improve the wear resistance of die casting dies. Final project report, January 1, 1995--April 30, 1997

    SciTech Connect

    Shivpuri, R.

    1997-09-18

    Die Casting dies are subject to severe service conditions during the die casting operation. While these severe conditions are necessary to achieve high production rates, they cause the dies which are commonly made of H13 die steel, to suffer frequent failures. The major die failure mechanisms are erosion or washout, Heat checking, soldering and corrosion. Due to their geometrical complexity, die casting dies are very expensive (some dies cost over a million dollars), and thus a large number of parts have to be produced by a die, to justify this cost and leverage the advantages of the die casting process (high production rates, low manpower costs). A potential increase in the die service life, thus has a significant impact on the economics of the die; casting operation. There are many ways to extend die life: developing new wear resistant die materials, developing new surface treatments including coatings, improving heat treatment of existing H13 dies, using better lubricants that can protect the die material, or modifying the die geometry and process parameters to reduce the intensity of wear. Of these the use of coatings to improve the wear resistance of the die surface has shown a lot of promise. Consequently, use of coatings in the die casting industry and their wide use to decrease die wear can improve significantly the productivity of shop operations resulting in large savings in material and energy usage.

  9. Elevated in vivo strontium-90 from nuclear weapons test fallout among cancer decedents: a case-control study of deciduous teeth.

    PubMed

    Mangano, Joseph J; Sherman, Janette D

    2011-01-01

    Risks to health from large-scale atmospheric nuclear weapons testing are still relatively unknown. A sample of 85,000 deciduous teeth collected from Americans born during the bomb-testing years assessed risk by in vivo measurement of residual strontium-90 (Sr-90) concentrations, using liquid scintillation spectrometry. The authors' analysis included 97 deciduous teeth from persons born between 1959 and 1961 who were diagrosed with cancer, and 194 teeth of matched controls. Average Sr-90 in teeth of persons who died of cancer was significantly greater than for controls (OR = 2.22; p < 0.04). This discovery suggests that many thousands have died or will die of cancer due to exposure to fallout, far more than previously believed. PMID:21319726

  10. Insights on TRP Channels from In Vivo Studies in Drosophila

    PubMed Central

    Minke, Baruch; Parnas, Moshe

    2007-01-01

    Transient receptor potential (TRP) channels mediate responses in a large variety of signaling mechanisms. Most studies on mammalian TRP channels rely on heterologous expression, but their relevance to in vivo tissues is not entirely clear. In contrast, Drosophila TRP and TRP-like (TRPL) channels allow direct analyses of in vivo function. In Drosophila photoreceptors, activation of TRP and TRPL is mediated via the phosphoinositide cascade, with both Ca2+ and diacylglycerol (DAG) essential for generating the light response. In tissue culture cells, TRPL channels are constitutively active, and lipid second messengers greatly facilitate this activity. Inhibition of phospholipase C (PLC) completely blocks lipid activation of TRPL, suggesting that lipid activation is mediated via PLC. In vivo studies in mutant Drosophila also reveal an acute requirement for lipid-producing enzyme, which may regulate PLC activity. Thus, PLC and its downstream second messengers, Ca2+ and DAG, constitute critical mediators of TRP/TRPL gating in vivo. PMID:16460287

  11. Imaging agents for in vivo magnetic resonance and scintigraphic imaging

    DOEpatents

    Engelstad, B.L.; Raymond, K.N.; Huberty, J.P.; White, D.L.

    1991-04-23

    Methods are provided for in vivo magnetic resonance imaging and/or scintigraphic imaging of a subject using chelated transition metal and lanthanide metal complexes. Novel ligands for these complexes are provided. No Drawings

  12. Novel Molecular and Nanosensors for In Vivo Sensing

    PubMed Central

    Eckert, Mark A.; Vu, Priscilla Q.; Zhang, Kaixiang; Kang, Dongku; Ali, M. Monsur; Xu, Chenjie; Zhao, Weian

    2013-01-01

    In vivo sensors are an emerging field with the potential to revolutionize our understanding of basic biology and our treatment of disease. In this review, we highlight recent advances in the fields of in vivo electrochemical, optical, and magnetic resonance biosensors with a focus on recent developments that have been validated in rodent models or human subjects. In addition, we discuss major challenges in the development and translation of in vivo biosensors and present potential solutions to these problems. The field of nanotechnology, in particular, has recently been instrumental in driving the field of in vivo sensors forward. We conclude with a discussion of emerging paradigms and techniques for the development of future biosensors. PMID:23946824

  13. The kinetics of ER fusion protein activation in vivo

    PubMed Central

    Wilson, Catherine H.; Gamper, Ivonne; Perfetto, Alessandra; Auw, Jeremy; Littlewood, Trevor D.; Evan, Gerard I.

    2014-01-01

    Reversibly switchable proteins are powerful tools with which to explore protein function in vitro and in vivo. For example, the activity of many proteins fused to the hormone-binding domain of the modified estrogen receptor (ERTAM) can be regulated by provision or removal of 4-hydroxytamoxifen (4-OHT). Despite the widespread use of ERTAM fusions in vivo, inadequate data are available as to the most efficacious routes for systemic tamoxifen delivery. In this study, we have used two well-characterised ERTAM fusion proteins, both reversibly activated by 4-OHT, to compare the effectiveness and kinetics of 4-OHT delivery in mice in vivo by either tamoxifen in food or by intraperitoneal injection. Our data indicate that dietary tamoxifen offers an effective, facile and ethically preferable means for long term activation of ERTAM fusion proteins in vivo. PMID:24662815

  14. A method to study in vivo stability of DNA nanostructures☆

    PubMed Central

    Surana, Sunaina; Bhatia, Dhiraj; Krishnan, Yamuna

    2013-01-01

    DNA nanostructures are rationally designed, synthetic, nanoscale assemblies obtained from one or more DNA sequences by their self-assembly. Due to the molecularly programmable as well as modular nature of DNA, such designer DNA architectures have great potential for in cellulo and in vivo applications. However, demonstrations of functionality in living systems necessitates a method to assess the in vivo stability of the relevant nanostructures. Here, we outline a method to quantitatively assay the stability and lifetime of various DNA nanostructures in vivo. This exploits the property of intact DNA nanostructures being uptaken by the coelomocytes of the multicellular model organism Caenorhabditis elegans. These studies reveal that the present fluorescence based assay in coelomocytes of C. elegans is an useful in vivo test bed for measuring DNA nanostructure stability. PMID:23623822

  15. The Expanding Toolbox of In Vivo Bioluminescent Imaging

    PubMed Central

    Xu, Tingting; Close, Dan; Handagama, Winode; Marr, Enolia; Sayler, Gary; Ripp, Steven

    2016-01-01

    In vivo bioluminescent imaging (BLI) permits the visualization of engineered bioluminescence from living cells and tissues to provide a unique perspective toward the understanding of biological processes as they occur within the framework of an authentic in vivo environment. The toolbox of in vivo BLI includes an inventory of luciferase compounds capable of generating bioluminescent light signals along with sophisticated and powerful instrumentation designed to detect and quantify these light signals non-invasively as they emit from the living subject. The information acquired reveals the dynamics of a wide range of biological functions that play key roles in the physiological and pathological control of disease and its therapeutic management. This mini review provides an overview of the tools and applications central to the evolution of in vivo BLI as a core technology in the preclinical imaging disciplines. PMID:27446798

  16. Cystoscopic optical coherence tomography for urinary bladder imaging in vivo

    NASA Astrophysics Data System (ADS)

    Wang, Z. G.; Adler, H.; Chan, D.; Jain, A.; Xie, H. K.; Wu, Z. L.; Pan, Y. T.

    2006-02-01

    This paper summarizes the development of new 2D MEMS mirrors and the pertinent modification to improve OCT endoscopic catheter packaging suitable for in vivo imaging diagnosis of bladder cancers. Comparative study of the newly developed endocopic OCT versus the bench-top OCT is presented. Results of in vivo OCT cystoscopy based on a porcine acute inflammation model are presented to compare time-domain OCT and spectral-domain OCT for in vivo imaging. In addition, results of spectral-domain Doppler OCT are presented to image blood flow in the lamina propria of the bladder. The results of our in vivo animal study using the presented OCT endoscope are discussed for potential problems in the future clinical applications.

  17. Imaging agents for in vivo magnetic resonance and scintigraphic imaging

    DOEpatents

    Engelstad, Barry L.; Raymond, Kenneth N.; Huberty, John P.; White, David L.

    1991-01-01

    Methods are provided for in vivo magnetic resonance imaging and/or scintigraphic imaging of a subject using chelated transition metal and lanthanide metal complexes. Novel ligands for these complexes are provided.

  18. Reactive polymer enables efficient in vivo bioorthogonal chemistry

    PubMed Central

    Devaraj, Neal K.; Thurber, Greg M.; Keliher, Edmund J.; Marinelli, Brett; Weissleder, Ralph

    2012-01-01

    There has been intense interest in the development of selective bioorthogonal reactions or “click” chemistry that can proceed in live animals. Until now however, most reactions still require vast surpluses of reactants because of steep temporal and spatial concentration gradients. Using computational modeling and design of pharmacokinetically optimized reactants, we have developed a predictable method for efficient in vivo click reactions. Specifically, we show that polymer modified tetrazines (PMT) are a key enabler for in vivo bioorthogonal chemistry based on the very fast and catalyst-free [4 + 2] tetrazine/trans-cyclooctene cycloaddition. Using fluorescent PMT for cellular resolution and 18F labeled PMT for whole animal imaging, we show that cancer cell epitopes can be easily reacted in vivo. This generic strategy should help guide the design of future chemistries and find widespread use for different in vivo bioorthogonal applications, particularly in the biomedical sciences. PMID:22411831

  19. In vivo spectral micro-imaging of tissue

    SciTech Connect

    Demos, Stavros G; Urayama, Shiro; Lin, Bevin; Saroufeem, Ramez; Ghobrial, Moussa

    2012-11-27

    In vivo endoscopic methods an apparatuses for implementation of fluorescence and autofluorescence microscopy, with and without the use of exogenous agents, effectively (with resolution sufficient to image nuclei) visualize and categorize various abnormal tissue forms.

  20. The Expanding Toolbox of In Vivo Bioluminescent Imaging.

    PubMed

    Xu, Tingting; Close, Dan; Handagama, Winode; Marr, Enolia; Sayler, Gary; Ripp, Steven

    2016-01-01

    In vivo bioluminescent imaging (BLI) permits the visualization of engineered bioluminescence from living cells and tissues to provide a unique perspective toward the understanding of biological processes as they occur within the framework of an authentic in vivo environment. The toolbox of in vivo BLI includes an inventory of luciferase compounds capable of generating bioluminescent light signals along with sophisticated and powerful instrumentation designed to detect and quantify these light signals non-invasively as they emit from the living subject. The information acquired reveals the dynamics of a wide range of biological functions that play key roles in the physiological and pathological control of disease and its therapeutic management. This mini review provides an overview of the tools and applications central to the evolution of in vivo BLI as a core technology in the preclinical imaging disciplines. PMID:27446798

  1. Portable optical fiber probe for in vivo brain temperature measurements.

    PubMed

    Musolino, Stefan; Schartner, Erik P; Tsiminis, Georgios; Salem, Abdallah; Monro, Tanya M; Hutchinson, Mark R

    2016-08-01

    This work reports on the development of an optical fiber based probe for in vivo measurements of brain temperature. By utilizing a thin layer of rare-earth doped tellurite glass on the tip of a conventional silica optical fiber a robust probe, suitable for long-term in vivo measurements of temperature can be fabricated. This probe can be interrogated using a portable optical measurement setup, allowing for measurements to be performed outside of standard optical laboratories. PMID:27570698

  2. Monitoring Retroviral RNA Dimerization In Vivo via Hammerhead Ribozyme Cleavage

    PubMed Central

    Pal, Bijay K.; Scherer, Lisa; Zelby, Laurie; Bertrand, Edouard; Rossi, John J.

    1998-01-01

    We have used a strategy for colocalization of Psi (Ψ)-tethered ribozymes and targets to demonstrate that Ψ sequences are capable of specific interaction in the cytoplasm of both packaging and nonpackaging cells. These results indicate that current in vitro dimerization models may have in vivo counterparts. The methodology used may be applied to further genetic analyses on Ψ domain interactions in vivo. PMID:9733882

  3. In vivo decomposition study of coated magnesium alloys

    NASA Astrophysics Data System (ADS)

    White, Desiree; Piersma, Tyler; Lecronier, David; Cheng, Xingguo; Rabago-Smith, Montserrat

    2010-04-01

    In the last decade, magnesium has resurged as an important biomaterial. Its mechanical properties are very similar to natural bone, and it degrades in vivo to non toxic substances. Unfortunately, corrosion of pure magnesium in vivo is rapid, thus coated alloys that decrease its corrosion could be used as implants in orthopedics. This presentation will describe the degradation results in cell cultures and in rats.

  4. Defining human mesenchymal stem cell efficacy in vivo.

    PubMed

    Bonfield, Tracey L; Nolan Koloze, Mary T; Lennon, Donald P; Caplan, Arnold I

    2010-01-01

    Allogeneic human mesenchymal stem cells (hMSCs) can suppress graft versus host disease (GvHD) and have profound anti-inflammatory and regenerative capacity in stroke, infarct, spinal cord injury, meniscus regeneration, tendinitis, acute renal failure, and heart disease in human and animal models of disease. There is significant clinical hMSC variability in efficacy and the ultimate response in vivo. The challenge in hMSC based therapy is defining the efficacy of hMSC in vivo. Models which may provide insight into hMSC bioactivity in vivo would provide a means to distinguish hMSCs for clinical utility. hMSC function has been described as both regenerative and trophic through the production of bioactive factors. The regenerative component involves the multi-potentiality of hMSC progenitor differentiation. The secreted factors generated by the hMSCs are milieu and injury specific providing unique niches for responses in vivo. These bioactive factors are anti-scarring, angiogenic, anti-apoptotic as well as regenerative. Further, from an immunological standpoint, hMSC's can avoid host immune response, providing xenographic applications. To study the in vivo immuno-regulatory effectiveness of hMSCs, we used the ovalbumin challenge model of acute asthma. This is a quick 3 week in vivo pulmonary inflammation model with readily accessible ways of measuring effectiveness of hMSCs. Our data show that there is a direct correlation between the traditional ceramic cube score to hMSCs attenuation of cellular recruitment due to ovalbumin challenge. The results from these studies verify the in vivo immuno-modulator effectiveness of hMSCs and support the potential use of the ovalbumin model as an in vivo model of hMSC potency and efficacy. Our data also support future directions toward exploring hMSCs as an alternative therapeutic for the treatment of airway inflammation associated with asthma. PMID:20974000

  5. Portable optical fiber probe for in vivo brain temperature measurements

    PubMed Central

    Musolino, Stefan; Schartner, Erik P.; Tsiminis, Georgios; Salem, Abdallah; Monro, Tanya M.; Hutchinson, Mark R.

    2016-01-01

    This work reports on the development of an optical fiber based probe for in vivo measurements of brain temperature. By utilizing a thin layer of rare-earth doped tellurite glass on the tip of a conventional silica optical fiber a robust probe, suitable for long-term in vivo measurements of temperature can be fabricated. This probe can be interrogated using a portable optical measurement setup, allowing for measurements to be performed outside of standard optical laboratories. PMID:27570698

  6. Vacuum die attach for integrated circuits

    DOEpatents

    Schmitt, Edward H.; Tuckerman, David B.

    1991-01-01

    A thin film eutectic bond for attaching an integrated circuit die to a circuit substrate is formed by coating at least one bonding surface on the die and substrate with an alloying metal, assembling the die and substrate under compression loading, and heating the assembly to an alloying temperature in a vacuum. A very thin bond, 10 microns or less, which is substantially void free, is produced. These bonds have high reliability, good heat and electrical conduction, and high temperature tolerance. The bonds are formed in a vacuum chamber, using a positioning and loading fixture to compression load the die, and an IR lamp or other heat source. For bonding a silicon die to a silicon substrate, a gold silicon alloy bond is used. Multiple dies can be bonded simultaneously. No scrubbing is required.

  7. Die singulation method and package formed thereby

    SciTech Connect

    Anderson, Robert C.; Shul, Randy J.; Clews, Peggy J.; Baker, Michael S.; De Boer, Maarten P.

    2012-08-07

    A method is disclosed for singulating die from a substrate having a sacrificial layer and one or more device layers, with a retainer being formed in the device layer(s) and anchored to the substrate. Deep Reactive Ion Etching (DRIE) etching of a trench through the substrate from the bottom side defines a shape for each die. A handle wafer is then attached to the bottom side of the substrate, and the sacrificial layer is etched to singulate the die and to form a frame from the retainer and the substrate. The frame and handle wafer, which retain the singulated die in place, can be attached together with a clamp or a clip and to form a package for the singulated die. One or more stops can be formed from the device layer(s) to limit a sliding motion of the singulated die.

  8. Reinraumtechnik für die Medizintechnik

    NASA Astrophysics Data System (ADS)

    Petek, Max; Jungbluth, Martin; Krampe, Erhard

    Die Reinraumtechnik ist heute ein unverzichtbarer Bestandteil bei der Fertigung von Produkten der Life Sciences, den Bereichen Pharma, Lebensmittel, Kosmetik und Medizintechnik. In Anbetracht der langen Historie der Medizintechnik ist sie jedoch eine sehr junge Disziplin. Die Bedeutung von Keimen und die richtige Einschätzung ihrer Größe wurden zwar sehr früh bereits durch Paracelsus erkannt, jedoch wurden daraus noch keine speziellen oder kontinuierlich umgesetzten Hygienevorschriften abgeleitet. Die erste bekannte technische Umsetzung von Hygieneempfehlungen geht auf den Franzosen François Nicolas Appert zurück, der eine aseptische Abfüllmethode für Lebensmittel entwickelte und diese 1810 veröffentlichte [1]. Die erste dokumentierte medizinische Umsetzung stellten Hygienevorschriften für Ärzte dar, die Ignaz Philipp Semmelweis nach 1847 in der Wiener Klinik für Geburtshilfe einführte [2].

  9. Vacuum die attach for integrated circuits

    DOEpatents

    Schmitt, E.H.; Tuckerman, D.B.

    1991-09-10

    A thin film eutectic bond for attaching an integrated circuit die to a circuit substrate is formed by coating at least one bonding surface on the die and substrate with an alloying metal, assembling the die and substrate under compression loading, and heating the assembly to an alloying temperature in a vacuum. A very thin bond, 10 microns or less, which is substantially void free, is produced. These bonds have high reliability, good heat and electrical conduction, and high temperature tolerance. The bonds are formed in a vacuum chamber, using a positioning and loading fixture to compression load the die, and an IR lamp or other heat source. For bonding a silicon die to a silicon substrate, a gold silicon alloy bond is used. Multiple dies can be bonded simultaneously. No scrubbing is required. 1 figure.

  10. Automatic in vivo portal dosimetry of all treatments

    NASA Astrophysics Data System (ADS)

    Olaciregui-Ruiz, I.; Rozendaal, R.; Mijnheer, B.; van Herk, M.; Mans, A.

    2013-11-01

    At our institution EPID (electronic portal imaging device) dosimetry is routinely applied to perform in vivo dose verification of all patient treatments with curative intent since January 2008. The major impediment of the method has been the amount of work required to produce and inspect the in vivo dosimetry reports (a time-consuming and labor-intensive process). In this paper we present an overview of the actions performed to implement an automated in vivo dosimetry solution clinically. We reimplemented the EPID dosimetry software and modified the acquisition software. Furthermore, we introduced new tools to periodically inspect the record-and-verify database and automatically run the EPID dosimetry software when needed. In 2012, 95% of our 3839 treatments scheduled for in vivo dosimetry were analyzed automatically (27 633 portal images of intensity-modulated radiotherapy (IMRT) fields, 5551 portal image data of VMAT arcs, and 2003 portal images of non-IMRT fields). The in vivo dosimetry verification results are available a few minutes after delivery and alerts are immediately raised when deviations outside tolerance levels are detected. After the clinical introduction of this automated solution, inspection of the detected deviations is the only remaining work. These newly developed tools are a major step forward towards full integration of in vivo EPID dosimetry in radiation oncology practice.

  11. Repeated swim stress alters brain benzodiazepine receptors measured in vivo

    SciTech Connect

    Weizman, R.; Weizman, A.; Kook, K.A.; Vocci, F.; Deutsch, S.I.; Paul, S.M.

    1989-06-01

    The effects of repeated swim stress on brain benzodiazepine receptors were examined in the mouse using both an in vivo and in vitro binding method. Specific in vivo binding of (/sup 3/H)Ro15-1788 to benzodiazepine receptors was decreased in the hippocampus, cerebral cortex, hypothalamus, midbrain and striatum after repeated swim stress (7 consecutive days of daily swim stress) when compared to nonstressed mice. In vivo benzodiazepine receptor binding was unaltered after repeated swim stress in the cerebellum and pons medulla. The stress-induced reduction in in vivo benzodiazepine receptor binding did not appear to be due to altered cerebral blood flow or to an alteration in benzodiazepine metabolism or biodistribution because there was no difference in (14C)iodoantipyrine distribution or whole brain concentrations of clonazepam after repeated swim stress. Saturation binding experiments revealed a change in both apparent maximal binding capacity and affinity after repeated swim stress. Moreover, a reduction in clonazepam's anticonvulsant potency was also observed after repeated swim stress (an increase in the ED50 dose for protection against pentylenetetrazol-induced seizures), although there was no difference in pentylenetetrazol-induced seizure threshold between the two groups. In contrast to the results obtained in vivo, no change in benzodiazepine receptor binding kinetics was observed using the in vitro binding method. These data suggest that environmental stress can alter the binding parameters of the benzodiazepine receptor and that the in vivo and in vitro binding methods can yield substantially different results.

  12. Automatic in vivo portal dosimetry of all treatments.

    PubMed

    Olaciregui-Ruiz, I; Rozendaal, R; Mijnheer, B; van Herk, M; Mans, A

    2013-11-21

    At our institution EPID (electronic portal imaging device) dosimetry is routinely applied to perform in vivo dose verification of all patient treatments with curative intent since January 2008. The major impediment of the method has been the amount of work required to produce and inspect the in vivo dosimetry reports (a time-consuming and labor-intensive process). In this paper we present an overview of the actions performed to implement an automated in vivo dosimetry solution clinically. We reimplemented the EPID dosimetry software and modified the acquisition software. Furthermore, we introduced new tools to periodically inspect the record-and-verify database and automatically run the EPID dosimetry software when needed. In 2012, 95% of our 3839 treatments scheduled for in vivo dosimetry were analyzed automatically (27,633 portal images of intensity-modulated radiotherapy (IMRT) fields, 5551 portal image data of VMAT arcs, and 2003 portal images of non-IMRT fields). The in vivo dosimetry verification results are available a few minutes after delivery and alerts are immediately raised when deviations outside tolerance levels are detected. After the clinical introduction of this automated solution, inspection of the detected deviations is the only remaining work. These newly developed tools are a major step forward towards full integration of in vivo EPID dosimetry in radiation oncology practice. PMID:24201085

  13. Should assisted dying be legalised?

    PubMed Central

    2014-01-01

    When an individual facing intractable pain is given an estimate of a few months to live, does hastening death become a viable and legitimate alternative for willing patients? Has the time come for physicians to do away with the traditional notion of healthcare as maintaining or improving physical and mental health, and instead accept their own limitations by facilitating death when requested? The Universities of Oxford and Cambridge held the 2013 Varsity Medical Debate on the motion “This House Would Legalise Assisted Dying”. This article summarises the key arguments developed over the course of the debate. We will explore how assisted dying can affect both the patient and doctor; the nature of consent and limits of autonomy; the effects on society; the viability of a proposed model; and, perhaps most importantly, the potential need for the practice within our current medico-legal framework. PMID:24423249

  14. In Vivo Osteogenic Differentiation of Human Dental Pulp Stem Cells Embedded in an Injectable In Vivo-Forming Hydrogel.

    PubMed

    Jang, Ja Yong; Park, Seung Hun; Park, Ji Hoon; Lee, Bo Keun; Yun, Jeong-Ho; Lee, Bong; Kim, Jae Ho; Min, Byoung Hyun; Kim, Moon Suk

    2016-08-01

    In this study, human dental pulp stem cells (hDPSCs) are examined as a cellular source for bone tissue engineering using an in vivo-forming hydrogel. The hDPSCs are easily harvested in large quantities from extracted teeth. The stemness of harvested hDPSCs indicates their relative tolerance to ex vivo manipulation in culture. The in vitro osteogenic differentiation of hDPSCs is characterized using Alizarin Red S (ARS), von Kossa (VK), and alkaline phosphatase (ALP) staining. The solution of hDPSCs and a methoxy polyethylene glycol-polycaprolactone block copolymer (PC) is easily prepared by simple mixing at room temperature and in no more than 10 s it forms in vivo hydrogels after subcutaneous injection into rats. In vivo osteogenic differentiation of hDPSCs in the in vivo-forming hydrogel is confirmed by micro-computed tomography (CT), histological staining, and gene expression. Micro-CT analysis shows evidence of significant tissue-engineered bone formation in hDPSCs-loaded hydrogel in the presence of osteogenic factors. Differentiated osteoblasts in in vivo-forming hydrogel are identified by ARS and VK staining and are found to exhibit characteristic expression of genes like osteonectin, osteopontin, and osteocalcin. In conclusion, hDPSCs embedded in an in vivo-forming hydrogel may provide benefits as a noninvasive formulation for bone tissue engineering applications. PMID:27074749

  15. In vivo exposure to hyperoxia increases airway responsiveness in rats. Demonstration in vivo and in vitro.

    PubMed

    Szarek, J L

    1989-10-01

    Studies regarding O2-induced lung injury have concentrated on damage to alveolar structures and pulmonary vasculature without consideration of alterations that may be occurring in airways. This study was undertaken to determine the effects of in vivo hyperoxic exposure on airway responses to excitatory stimuli in intact, anesthetized rats and in intrapulmonary bronchi isolated from hyperoxia-exposed rats. Using lung conductance (G1) as an index of bronchoconstriction, intravenously administered 5-hydroxytryptamine (5HT) elicited greater bronchoconstrictor responses in anesthetized, mechanically ventilated rats that had been exposed to 85% O2 for 7 days rather than to air. Further, airways of hyperoxia-exposed rats were more sensitive to the effects of intravenously administered 5HT as evidenced by the lower log dose of 5HT required to decrease G1 30%. Cylindrical segments of intrapulmonary bronchi isolated from hyperoxia-exposed rats were more responsive to the contractile effects of 5HT and electrical field stimulation. However, no differences in responsiveness to bethanechol or KCl were observed between the two groups. The log concentration of 5HT and the log frequency of electrical field stimulation that elicited half-maximal responses were smaller in bronchi isolated from hyperoxia-exposed animals, indicating an increase in sensitivity of the airways to these stimuli. These results suggest that prolonged exposure to greater than ambient levels of O2 can alter airway function; however, the mechanism responsible for these changes remains to be determined. PMID:2802379

  16. Prediction of Part Distortion in Die Casting

    SciTech Connect

    R. Allen Miller

    2005-03-30

    The die casting process is one of the net shape manufacturing techniques and is widely used to produce high production castings with tight tolerances for many industries. An understanding of the stress distribution and the deformation pattern of parts produced by die casting will result in less deviation from the part design specification, a better die design and eventually more productivity and cost savings. This report presents methods that can be used to simulate the die casting process in order to predict the deformation and stresses in the produced part and assesses the degree to which distortion modeling is practical for die casting at the current time. A coupled thermal-mechanical finite elements model was used to simulate the die casting process. The simulation models the effect of thermal and mechanical interaction between the casting and the die. It also includes the temperature dependant material properties of the casting. Based on a designed experiment, a sensitivity analysis was conducted on the model to investigate the effect of key factors. These factors include the casting material model, material properties and thermal interaction between casting and dies. To verify the casting distortion predictions, it was compared against the measured dimensions of produced parts. The comparison included dimensions along and across the parting plane and the flatness of one surface.

  17. Student Nurses' Perception of Death and Dying

    ERIC Educational Resources Information Center

    Niederriter, Joan E.

    2009-01-01

    Student nurses are involved in caring for patients who are actively dying or who have been told they have a terminal illness and are faced with the process of dying. Students encounter these patients in hospitals, nursing homes, at home or in hospice care settings. According to Robinson (2004), "nurses are the healthcare providers that are most…

  18. The Right To Die. Public Talk Series.

    ERIC Educational Resources Information Center

    Pasquerella, Lynn

    This program guide on the right to die provides policy issue information where ethical concerns have a prominent place. Three positions about the right to die are presented: (1) mercy killing and assisted suicide should be legally permitted in certain cases; (2) legal status should be given to living wills and other advance directives that would…

  19. Die Evolution der Religiosität

    NASA Astrophysics Data System (ADS)

    Voland, Eckart

    Ein konsequent darwinischer Blick auf den Menschen bedeutet, auch im Denken, Fühlen und Handeln biologische Anpassungsgeschichte zu suchen, denn auch die psychischen und mentalen Eigenheiten des Homo sapiens unterliegen der natürlichen Selektion. Lässt sich die religiöse Lebenspraxis von Menschen daher auch aus einer Fitnessperspektive betrachten?

  20. Energy Consumption of Die Casting Operations

    SciTech Connect

    Jerald Brevick; clark Mount-Campbell; Carroll Mobley

    2004-03-15

    Molten metal processing is inherently energy intensive and roughly 25% of the cost of die-cast products can be traced to some form of energy consumption [1]. The obvious major energy requirements are for melting and holding molten alloy in preparation for casting. The proper selection and maintenance of melting and holding equipment are clearly important factors in minimizing energy consumption in die-casting operations [2]. In addition to energy consumption, furnace selection also influences metal loss due to oxidation, metal quality, and maintenance requirements. Other important factors influencing energy consumption in a die-casting facility include geographic location, alloy(s) cast, starting form of alloy (solid or liquid), overall process flow, casting yield, scrap rate, cycle times, number of shifts per day, days of operation per month, type and size of die-casting form of alloy (solid or liquid), overall process flow, casting yield, scrap rate, cycle times, number of shifts per day, days of operation per month, type and size of die-casting machine, related equipment (robots, trim presses), and downstream processing (machining, plating, assembly, etc.). Each of these factors also may influence the casting quality and productivity of a die-casting enterprise. In a die-casting enterprise, decisions regarding these issues are made frequently and are based on a large number of factors. Therefore, it is not surprising that energy consumption can vary significantly from one die-casting enterprise to the next, and within a single enterprise as function of time.

  1. Assisted dying: a review of international legislation.

    PubMed

    Field, Charlotte; Curtice, Martin

    2009-05-01

    The issue of assisted dying in the UK is increasingly receiving media and academic journal attention. Such reporting often cites, but in little depth, existing legislation in other countries. Such international legislation may also shape future UK assisted dying legislation. PMID:19451872

  2. Apparatus for restraining and transporting dies

    DOEpatents

    Allison, James W.; LaBarre, Timothy L.

    1994-01-01

    Apparatus for restraining and transporting dies in punch press operations is provided. A floatation platen for supporting a die on the platen's upper surface has a plurality of recessed gas exhaust ports on the platen's lower surface. A source of pressurized gas delivers gas to a platen manifold, for delivery to orifices located in the gas exhaust ports. The flow of gas is controlled by a first valve adjacent the gas source and a second valve adjacent the manifold, with the second valve being used to control the gas flow during movement of the die. In this fashion, a die may be moved on a cushion of air from one workstation to a selected second workstation. A moveable hydraulically operated restraining fixture is also provided, for clamping the die in position during the compacting phase, and for releasing the die after completion of the compacting phase by releasing the hydraulic pressure on the restraining fixture. When pressure in the hydraulic cylinders on the restraining fixture is reversed, the restraining fixture will retract so that there is no contact between the die and the restraining fixture, thereby allowing the die to be removed from a first workstation and moved to a second selected workstation.

  3. In Vivo Application of Photocleavable Protein Interaction Reporter Technology

    PubMed Central

    Yang, Li; Zheng, Chunxiang; Weisbrod, Chad R.; Tang, Xiaoting; Munske, Gerhard R.; Hoopmann, Michael R.; Eng, Jimmy K.; Bruce, James E.

    2012-01-01

    Summary In vivo protein structures and protein-protein interactions are critical to the function of proteins in biological systems. As a complementary approach to traditional protein interaction identification methods, cross-linking strategies are beginning to provide additional data on protein and protein complex topological features. Previously, photocleavable protein interaction reporter (pcPIR) technology was demonstrated by cross-linking pure proteins and protein complexes and the use of ultraviolet light to cleave or release cross-linked peptides to enable identification. In the present report, the pcPIR strategy is applied to E. coli cells and in vivo protein interactions and topologies are measured. More than 1600 labeled peptides from E. coli were identified, indicating many protein sites react with pcPIR in vivo. From those labeled sites, 53 in vivo inter-cross-linked peptide pairs were identified and manually validated. Approximately half of the interactions have been reported using other techniques, although detailed structures exist for very few. Three proteins or protein complexes with detailed crystallography structures are compared to the cross-linking results obtained from in vivo application of pcPIR technology. PMID:22168182

  4. In Vitro and In Vivo Neurotoxicity of Prion Protein Oligomers

    PubMed Central

    Simoneau, Steve; Rezaei, Human; Salès, Nicole; Kaiser-Schulz, Gunnar; Lefebvre-Roque, Maxime; Vidal, Catherine; Fournier, Jean-Guy; Comte, Julien; Wopfner, Franziska; Grosclaude, Jeanne; Schätzl, Hermann; Lasmézas, Corinne Ida

    2007-01-01

    The mechanisms underlying prion-linked neurodegeneration remain to be elucidated, despite several recent advances in this field. Herein, we show that soluble, low molecular weight oligomers of the full-length prion protein (PrP), which possess characteristics of PrP to PrPsc conversion intermediates such as partial protease resistance, are neurotoxic in vitro on primary cultures of neurons and in vivo after subcortical stereotaxic injection. Monomeric PrP was not toxic. Insoluble, fibrillar forms of PrP exhibited no toxicity in vitro and were less toxic than their oligomeric counterparts in vivo. The toxicity was independent of PrP expression in the neurons both in vitro and in vivo for the PrP oligomers and in vivo for the PrP fibrils. Rescue experiments with antibodies showed that the exposure of the hydrophobic stretch of PrP at the oligomeric surface was necessary for toxicity. This study identifies toxic PrP species in vivo. It shows that PrP-induced neurodegeneration shares common mechanisms with other brain amyloidoses like Alzheimer disease and opens new avenues for neuroprotective intervention strategies of prion diseases targeting PrP oligomers. PMID:17784787

  5. Endocavitary in vivo Dosimetry for IMRT Treatments of Gynecologic Tumors

    SciTech Connect

    Cilla, Savino; Macchia, Gabriella; Digesu, Cinzia; Deodato, Francesco; Sabatino, Domenico; Morganti, Alessio G.; Piermattei, Angelo

    2011-01-01

    The accuracy and reproducibility of endometrial carcinoma treatment with intensity-modulated radiotherapy (IMRT) was assessed by means of in vivo dosimetry. Six patients who had previously undergone radical hysterectomy for endometrial carcinoma were treated with IMRT using a vaginal applicator with radio-opaque fiducial markers. An ion-chamber inserted into the applicator supplied an endocavitary in vivo dosimetry for quality assurance purposes. The ratio R = D/D{sub TPS} between the in vivo measured dose D and the predicted dose by the treatment planning system D{sub TPS} was determined for every fraction of the treatment. Results showed that 90% and 100% of the ratios resulted equal to 1 within 5% and 10%, respectively. The mean value of the ratios distribution for the 6 patients was R = 0.995 and the SD = 0.034. The ratio R* between the measured and predicted total doses for each patient was near to 1, within 2%. The dosimetric results suggest that the use of a vaginal applicator in an image-guided approach could make the interfractions target position stable and reproducible, allowing a safe use of the IMRT technique in the treatment of postoperative vaginal vault. In vivo dosimetry may supply useful information about the discrimination of random vs. systematic errors. The workload is minimum and this in vivo dosimetry can be applied also in the clinical routine.

  6. Transcriptional complexity of vaccinia virus in vivo and in vitro.

    PubMed Central

    Paoletti, E; Grady, L J

    1977-01-01

    The transcriptional complexity of vaccinia virus both in vivo and in vitro has been measured by using DNA:RNA hybridization with RNA in excess. In vivo, "early" or prereplicative RNA was found to saturate at 25% or one-half of the viral genome. "Late" or postreplicative RNA from infected HeLa cells saturated at 52% or essentially the entire genome. This well-regulated transcriptional pattern of the virus in vivo was not maintained in vitro. In a number of experiments a range of saturation values from 40 to 50% was obtained for in vitro synthesized RNA. The complexity of polyadenylated and non-polyadenylated RNA, as well as total purified 8 to 12S RNA released from the virus, was indistinguishable from purified high-molecular-weight virion-associated RNA with a sedimentation value of greater than 20S and equivalent to total in vitro synthesized RNA. No additional hybrid formation was observed in experiments in which total in vitro RNA and late in vivo RNA from infected HeLa cells were combined, suggesting that the virus does not transcribe in vitro DNA sequences that are not also transcribed during productive infection. Approximately 15% complementary RNA was detected when radiolabeled total in vitro RNA was allowed to reanneal with late in vivo RNA, while as much as 8% of the in vitro synthesized RNA was found to be complementary. PMID:894791

  7. In vivo bubble nucleation probability in sheep brain tissue.

    PubMed

    Gateau, J; Aubry, J-F; Chauvet, D; Boch, A-L; Fink, M; Tanter, M

    2011-11-21

    Gas nuclei exist naturally in living bodies. Their activation initiates cavitation activity, and is possible using short ultrasonic excitations of high amplitude. However, little is known about the nuclei population in vivo, and therefore about the rarefaction pressure required to form bubbles in tissue. A novel method dedicated to in vivo investigations was used here that combines passive and active cavitation detection with a multi-element linear ultrasound probe (4-7 MHz). Experiments were performed in vivo on the brain of trepanated sheep. Bubble nucleation was induced using a focused single-element transducer (central frequency 660 kHz, f-number = 1) driven by a high power (up to 5 kW) electric burst of two cycles. Successive passive recording and ultrafast active imaging were shown to allow detection of a single nucleation event in brain tissue in vivo. Experiments carried out on eight sheep allowed statistical studies of the bubble nucleation process. The nucleation probability was evaluated as a function of the peak negative pressure. No nucleation event could be detected with a peak negative pressure weaker than -12.7 MPa, i.e. one order of magnitude higher than the recommendations based on the mechanical index. Below this threshold, bubble nucleation in vivo in brain tissues is a random phenomenon. PMID:22015981

  8. Reproduction of in vivo motion using a parallel robot.

    PubMed

    Howard, Ryan A; Rosvold, Joshua M; Darcy, Shon P; Corr, David T; Shrive, Nigel G; Tapper, Janet E; Ronsky, Janet L; Beveridge, Jillian E; Marchuk, Linda L; Frank, Cyril B

    2007-10-01

    Although alterations in knee joint loading resulting from injury have been shown to influence the development of osteoarthritis, actual in vivo loading conditions of the joint remain unknown. A method for determining in vivo ligament loads by reproducing joint specific in vivo kinematics using a robotic testing apparatus is described. The in vivo kinematics of the ovine stifle joint during walking were measured with 3D optical motion analysis using markers rigidly affixed to the tibia and femur. An additional independent single degree of freedom measuring device was also used to record a measure of motion. Following sacrifice, the joint was mounted in a robotic/universal force sensor test apparatus and referenced using a coordinate measuring machine. A parallel robot configuration was chosen over the conventional serial manipulator because of its greater accuracy and stiffness. Median normal gait kinematics were applied to the joint and the resulting accuracy compared. The mean error in reproduction as determined by the motion analysis system varied between 0.06 mm and 0.67 mm and 0.07 deg and 0.74 deg for the two individual tests. The mean error measured by the independent device was found to be 0.07 mm and 0.83 mm for the two experiments, respectively. This study demonstrates the ability of this system to reproduce in vivo kinematics of the ovine stifle joint in vitro. The importance of system stiffness is discussed to ensure accurate reproduction of joint motion. PMID:17887900

  9. Systemic inflammation regulates microglial responses to tissue damage in vivo.

    PubMed

    Gyoneva, Stefka; Davalos, Dimitrios; Biswas, Dipankar; Swanger, Sharon A; Garnier-Amblard, Ethel; Loth, Francis; Akassoglou, Katerina; Traynelis, Stephen F

    2014-08-01

    Microglia, the resident immune cells of the central nervous system, exist in either a "resting" state associated with physiological tissue surveillance or an "activated" state in neuroinflammation. We recently showed that ATP is the primary chemoattractor to tissue damage in vivo and elicits opposite effects on the motility of activated microglia in vitro through activation of adenosine A2A receptors. However, whether systemic inflammation affects microglial responses to tissue damage in vivo remains largely unknown. Using in vivo two-photon imaging of mice, we show that injection of lipopolysaccharide (LPS) at levels that can produce both clear neuroinflammation and some features of sepsis significantly reduced the rate of microglial response to laser-induced ablation injury in vivo. Under proinflammatory conditions, microglial processes initially retracted from the ablation site, but subsequently moved toward and engulfed the damaged area. Analyzing the process dynamics in 3D cultures of primary microglia indicated that only A2A , but not A1 or A3 receptors, mediate process retraction in LPS-activated microglia. The A2A receptor antagonists caffeine and preladenant reduced adenosine-mediated process retraction in activated microglia in vitro. Finally, administration of preladenant before induction of laser ablation in vivo accelerated the microglial response to injury following systemic inflammation. The regulation of rapid microglial responses to sites of injury by A2A receptors could have implications for their ability to respond to the neuronal death occurring under conditions of neuroinflammation in neurodegenerative disorders. PMID:24807189

  10. Analysis of the structure of human telomerase RNA in vivo

    PubMed Central

    Antal, Mária; Boros, Éva; Solymosy, Ferenc; Kiss, Tamás

    2002-01-01

    Telomerase is a ribonucleoprotein reverse transcriptase that synthesises telomeric DNA. The RNA component of telomerase acts as a template for telomere synthesis and binds the reverse transcriptase. In this study, we have performed in vivo and in vitro structural analyses of human telomerase RNA (hTR). In vivo mapping experiments showed that the 5′-terminal template domain of hTR folds into a long hairpin structure, in which the template sequence occupies a readily accessible position. Intriguingly, neither in vivo nor in vitro mapping of hTR confirmed formation of a stable ‘pseudoknot’ helix, suggesting that this functionally essential long range interaction is formed only temporarily. In vitro control mappings demonstrated that the 5′-terminal template domain of hTR cannot fold correctly in the absence of cellular protein factors. The 3′-terminal domain of hTR, both in vivo and in vitro, folds into the previously predicted box H/ACA snoRNA-like ‘hairpin–hinge–hairpin–tail’ structure. Finally, comparison of the in vivo and in vitro modification patterns of hTR revealed several regions that might be directly involved in binding of telomerase reverse transcriptase or other telomerase proteins. PMID:11842102

  11. Pharmacokinetic Modeling of an Induction Regimen for In Vivo Combined Testing of Novel Drugs against Pediatric Acute Lymphoblastic Leukemia Xenografts

    PubMed Central

    Kang, Min H.; Liem, Natalia L. M.; Carol, Hernan; Boehm, Ingrid; Groepper, Daniel; Reynolds, C. Patrick; Stewart, Clinton F.; Lock, Richard B.

    2012-01-01

    Current regimens for induction therapy of pediatric acute lymphoblastic leukemia (ALL), or for re-induction post relapse, use a combination of vincristine (VCR), a glucocorticoid, and l-asparaginase (ASP) with or without an anthracycline. With cure rates now approximately 80%, robust pre-clinical models are necessary to prioritize active new drugs for clinical trials in relapsed/refractory patients, and the ability of these models to predict synergy/antagonism with established therapy is an essential attribute. In this study, we report optimization of an induction-type regimen by combining VCR, dexamethasone (DEX) and ASP (VXL) against ALL xenograft models established from patient biopsies in immune-deficient mice. We demonstrate that the VXL combination was synergistic in vitro against leukemia cell lines as well as in vivo against ALL xenografts. In vivo, VXL treatment caused delays in progression of individual xenografts ranging from 22 to >146 days. The median progression delay of xenografts derived from long-term surviving patients was 2-fold greater than that of xenografts derived from patients who died of their disease. Pharmacokinetic analysis revealed that systemic DEX exposure in mice increased 2-fold when administered in combination with VCR and ASP, consistent with clinical findings, which may contribute to the observed synergy between the 3 drugs. Finally, as proof-of-principle we tested the in vivo efficacy of combining VXL with either the Bcl-2/Bcl-xL/Bcl-w inhibitor, ABT-737, or arsenic trioxide to provide evidence of a robust in vivo platform to prioritize new drugs for clinical trials in children with relapsed/refractory ALL. PMID:22479469

  12. [Right to die with dignity?].

    PubMed

    Ruiz, Alvaro

    2008-06-01

    The right to die with dignity is an ill-defined concept, with multiple, often inappropriate, interpretations. The current proposition is that the physician take full responsibility for protecting the patients rights, for ensuring a rational use of resources and for overseeing the decision-making process such that the information is adequate and the steps proportioned. This responsibility extends not only to the health status of the patient situation, to the patients prognosis, and to his/her expectations and wishes, but also to the benefits foreseen and to the cost-benefit ratio. Emphasis is placed on two aspects of this relationship. First, dignity can be interpreted in many ways and sometimes, in the name of dignity, the patient is exposed (or exposes him/herself) to suffering, pain and complications that can be avoided. Second, when no reasonable probability of survival is present and a better quality of life is impossible, efforts are better redirected to offering a better quality of death. PMID:18719721

  13. Dying to go to school.

    PubMed

    Shearar, A

    1997-01-01

    In southern Sudan, the recent war sparked a mass migration of boys aged 5-18 who traveled through perilous terrain to the borders of Ethiopia in search of promised security and schooling. Thousands of these children died from hardships suffered in the wild, from the inadvertent or deliberate attacks of warriors, from hunger, from thirst, or from disease. Those who endured the hardships and arrived at their destination were shocked to find only military training centers or rudimentary schooling for those who were too weak or too young to be trained for battle. Eventually, the refugees were forced back into Sudan, and today about 20,000 of these unaccompanied children are displaced. Several nongovernmental organizations are running family reunification projects, and follow-up activities carried out among the reunified children reveal that they carry the scars of their trauma. The drawings the children produce as art therapy usually include the image of a young child holding a school book. While these children have a constant struggle with their memories, those who have not returned home or who have lost their families to war or disease are even more forlorn. As food rations have been halved in the refugee camps, many children are pursuing any possible alternative living arrangement. Some attempt to travel to South Africa or Egypt and some join an army in an effort to overcome the desperation of their present situation. PMID:12294200

  14. Quantitating intracellular oxygen tension in vivo by phosphorescence lifetime measurement

    PubMed Central

    Hirakawa, Yosuke; Yoshihara, Toshitada; Kamiya, Mako; Mimura, Imari; Fujikura, Daichi; Masuda, Tsuyoshi; Kikuchi, Ryohei; Takahashi, Ippei; Urano, Yasuteru; Tobita, Seiji; Nangaku, Masaomi

    2015-01-01

    Hypoxia appears to have an important role in pathological conditions in many organs such as kidney; however, a method to quantify intracellular oxygen tension in vivo has not been well established. In this study, we established an optical method to quantify oxygen tension in mice kidneys using a cationic lipophilic phosphorescence probe, BTPDM1, which has an intracellular oxygen concentration-sensitive phosphorescence lifetime. Since this probe is distributed inside the tubular cells of the mice kidney, we succeeded in detecting acute renal hypoxic conditions and chronic kidney disease. This technique enabled us to estimate intracellular partial pressures of oxygen in vivo by extrapolating the calibration curve generated from cultured tubular cells. Since intracellular oxygen tension is directly related to cellular hypoxic reactions, such as the activation of hypoxia-inducible factors, our method will shed new light on hypoxia research in vivo. PMID:26644023

  15. Delivery of Therapeutic RNAs Into Target Cells IN VIVO

    NASA Astrophysics Data System (ADS)

    Ng, Mei Ying; Hagen, Thilo

    2014-02-01

    RNA-based therapy is one of the most promising approaches to treat human diseases. Specifically, the use of short interfering RNA (siRNA) siRNA and microRNA (miRNA) mimics for in vivo RNA interference has immense potential as it directly lowers the expression of the therapeutic target protein. However, there are a number of major roadblocks to the successful implementation of siRNA and other RNA based therapies in the clinic. These include the instability of RNAs in vivo and the difficulty to efficiently deliver the RNA into the target cells. Hence, various innovative approaches have been taken over the years to develop effective RNA delivery methods. These methods include liposome-, polymeric nanoparticle- and peptide-mediated cellular delivery. In a recent innovative study, bioengineered bacterial outer membrane vesicles were used as vehicles for effective delivery of siRNA into cells in vivo.

  16. In vitro-In vivo Correlation: Perspectives on Model Development

    PubMed Central

    Lu, Ying; Kim, Sungwon; Park, Kinam

    2011-01-01

    In vitro – in vivo correlation (IVIVC) allows prediction of the in vivo performance of a drug based on the in vitro drug release profiles. To develop an effective IVIVC, the physicochemical and biopharmaceutical properties of the drug as well as the physiological environment in the body must be taken into consideration. Key factors include drug solubility, pKa, drug permeability, octanol-water partition coefficient and pH of environment. In general, construction of an IVIVC involves three stages of mathematical manipulation: construct a functional relationship between input (in vitro dissolution) and output (in vivo dissolution); establish a structural relationship using data collected; parameterize the unknowns in the structural model. Some key mathematical relationships used in IVIVC development are presented. The establishment of an effective IVIVC has important implications in quality control and regulatory compliance. PMID:21237256

  17. Progress Toward In Vivo Use of siRNAs-II

    PubMed Central

    Rettig, Garrett R; Behlke, Mark A

    2012-01-01

    RNA interference (RNAi) has been extensively employed for in vivo research since its use was first demonstrated in mammalian cells 10 years ago. Design rules have improved, and it is now routinely possible to obtain reagents that suppress expression of any gene desired. At the same time, increased understanding of the molecular basis of unwanted side effects has led to the development of chemical modification strategies that mitigate these concerns. Delivery remains the single greatest hurdle to widespread adoption of in vivo RNAi methods. However, exciting advances have been made and new delivery systems under development may help to overcome these barriers. This review discusses advances in RNAi biochemistry and biology that impact in vivo use and provides an overview of select publications that demonstrate interesting applications of these principles. Emphasis is placed on work with synthetic, small interfering RNAs (siRNAs) published since the first installment of this review which appeared in 2006. PMID:22186795

  18. A biomagnetic system for in vivo cancer imaging

    PubMed Central

    Flynn, E R; Bryant, H C

    2007-01-01

    An array of highly sensitive biomagnetic sensors of the superconducting quantum interference detector (SQUID) type can identify disease in vivo by detecting and imaging microscopic amounts of nanoparticles. We describe in detail procedures and parameters necessary for implementation of in vivo detection through the use of antibody-labelled magnetic nanoparticles as well as methods of determining magnetic nanoparticle properties. We discuss the weak field magnetic sensor SQUID system, the method of generating the magnetic polarization pulse to align the magnetic moments of the nanoparticles, and the measurement techniques to measure their magnetic remanence fields following this pulsed field. We compare these results to theoretical calculations and predict optimal properties of nanoparticles for in vivo detection. PMID:15798322

  19. In vivo modulation of endothelial polarization by Apelin receptor signalling

    PubMed Central

    Kwon, Hyouk-Bum; Wang, Shengpeng; Helker, Christian S. M.; Rasouli, S. Javad; Maischein, Hans-Martin; Offermanns, Stefan; Herzog, Wiebke; Stainier, Didier Y. R.

    2016-01-01

    Endothelial cells (ECs) respond to shear stress by aligning in the direction of flow. However, how ECs respond to flow in complex in vivo environments is less clear. Here we describe an endothelial-specific transgenic zebrafish line, whereby the Golgi apparatus is labelled to allow for in vivo analysis of endothelial polarization. We find that most ECs polarize within 4.5 h after the onset of vigorous blood flow and, by manipulating cardiac function, observe that flow-induced EC polarization is a dynamic and reversible process. Based on its role in EC migration, we analyse the role of Apelin signalling in EC polarization and find that it is critical for this process. Knocking down Apelin receptor function in human primary ECs also affects their polarization. Our study provides new tools to analyse the mechanisms of EC polarization in vivo and reveals an important role in this process for a signalling pathway implicated in cardiovascular disease. PMID:27248505

  20. In vivo modulation of endothelial polarization by Apelin receptor signalling.

    PubMed

    Kwon, Hyouk-Bum; Wang, Shengpeng; Helker, Christian S M; Rasouli, S Javad; Maischein, Hans-Martin; Offermanns, Stefan; Herzog, Wiebke; Stainier, Didier Y R

    2016-01-01

    Endothelial cells (ECs) respond to shear stress by aligning in the direction of flow. However, how ECs respond to flow in complex in vivo environments is less clear. Here we describe an endothelial-specific transgenic zebrafish line, whereby the Golgi apparatus is labelled to allow for in vivo analysis of endothelial polarization. We find that most ECs polarize within 4.5 h after the onset of vigorous blood flow and, by manipulating cardiac function, observe that flow-induced EC polarization is a dynamic and reversible process. Based on its role in EC migration, we analyse the role of Apelin signalling in EC polarization and find that it is critical for this process. Knocking down Apelin receptor function in human primary ECs also affects their polarization. Our study provides new tools to analyse the mechanisms of EC polarization in vivo and reveals an important role in this process for a signalling pathway implicated in cardiovascular disease. PMID:27248505

  1. In Vivo Reprogramming for Brain and Spinal Cord Repair.

    PubMed

    Chen, Gong; Wernig, Marius; Berninger, Benedikt; Nakafuku, Masato; Parmar, Malin; Zhang, Chun-Li

    2015-01-01

    Cell reprogramming technologies have enabled the generation of various specific cell types including neurons from readily accessible patient cells, such as skin fibroblasts, providing an intriguing novel cell source for autologous cell transplantation. However, cell transplantation faces several difficult hurdles such as cell production and purification, long-term survival, and functional integration after transplantation. Recently, in vivo reprogramming, which makes use of endogenous cells for regeneration purpose, emerged as a new approach to circumvent cell transplantation. There has been evidence for in vivo reprogramming in the mouse pancreas, heart, and brain and spinal cord with various degrees of success. This mini review summarizes the latest developments presented in the first symposium on in vivo reprogramming glial cells into functional neurons in the brain and spinal cord, held at the 2014 annual meeting of the Society for Neuroscience in Washington, DC. PMID:26730402

  2. Handheld multispectral fluorescence lifetime imaging system for in vivo applications.

    PubMed

    Cheng, Shuna; Cuenca, Rodrigo M; Liu, Boang; Malik, Bilal H; Jabbour, Joey M; Maitland, Kristen C; Wright, John; Cheng, Yi-Shing Lisa; Jo, Javier A

    2014-03-01

    There is an increasing interest in the application of fluorescence lifetime imaging (FLIM) for medical diagnosis. Central to the clinical translation of FLIM technology is the development of compact and high-speed clinically compatible systems. We present a handheld probe design consisting of a small maneuverable box fitted with a rigid endoscope, capable of continuous lifetime imaging at multiple emission bands simultaneously. The system was characterized using standard fluorescent dyes. The performance was then further demonstrated by imaging a hamster cheek pouch in vivo, and oral mucosa tissue both ex vivo and in vivo, all using safe and permissible exposure levels. Such a design can greatly facilitate the evaluation of FLIM for oral cancer imaging in vivo. PMID:24688824

  3. RNA circularization strategies in vivo and in vitro

    PubMed Central

    Petkovic, Sonja; Müller, Sabine

    2015-01-01

    In the plenitude of naturally occurring RNAs, circular RNAs (circRNAs) and their biological role were underestimated for years. However, circRNAs are ubiquitous in all domains of life, including eukaryotes, archaea, bacteria and viruses, where they can fulfill diverse biological functions. Some of those functions, as for example playing a role in the life cycle of viral and viroid genomes or in the maturation of tRNA genes, have been elucidated; other putative functions still remain elusive. Due to the resistance to exonucleases, circRNAs are promising tools for in vivo application as aptamers, trans-cleaving ribozymes or siRNAs. How are circRNAs generated in vivo and what approaches do exist to produce ring-shaped RNAs in vitro? In this review we illustrate the occurrence and mechanisms of RNA circularization in vivo, survey methods for the generation of circRNA in vitro and provide appropriate protocols. PMID:25662225

  4. In vivo genomic footprint of a yeast centromere.

    PubMed Central

    Densmore, L; Payne, W E; Fitzgerald-Hayes, M

    1991-01-01

    We have used in vivo genomic footprinting to investigate the protein-DNA interactions within the conserved DNA elements (CDEI, CDEII, and CDEIII) in the centromere from chromosome III of the yeast Saccharomyces cerevisiae. The in vivo footprint pattern obtained from wild-type cells shows that some guanines within the centromere DNA are protected from methylation by dimethyl sulfate. These results are consistent with studies demonstrating that yeast cells contain sequence-specific centromere DNA-binding proteins. Our in vivo experiments on chromosomes with mutant centromeres show that some mutations which affect chromosome segregation also alter the footprint pattern caused by proteins bound to the centromere DNA. The results of this study provide the first fine-structure map of proteins bound to centromere DNA in living yeast cells and suggest a direct correlation between these protein-DNA interactions and centromere function. Images PMID:1986217

  5. Photoacoustic molecular imaging for in vivo liver iron quantitation

    NASA Astrophysics Data System (ADS)

    Maccarinelli, Federica; Carmona, Fernando; Regoni, Maria; Arosio, Paolo

    2016-05-01

    A recent study showed that ferritin is a suitable endogenous contrast agent for photoacoustic molecular imaging in cultured mammalian cells. We have therefore tested whether this imaging technique can be used for in vivo quantification of iron in mouse livers. To verify this hypothesis, we used multispectral optoacoustic tomography (MSOT) to image albino CD1 mice before and after experimental iron loading. Postmortem assays showed that the iron treatment caused a 15-fold increase in liver iron and a 40-fold increase in liver ferritin levels, while in vivo longitudinal analysis using MSOT revealed just a 1.6-fold increase in the ferritin/iron photoacoustic signal in the same animals. We conclude that MSOT can monitor changes in ferritin/iron levels in vivo, but its sensitivity is much lower than that of ex vivo iron assays.

  6. In vivo acoustic and photoacoustic focusing of circulating cells

    NASA Astrophysics Data System (ADS)

    Galanzha, Ekaterina I.; Viegas, Mark G.; Malinsky, Taras I.; Melerzanov, Alexander V.; Juratli, Mazen A.; Sarimollaoglu, Mustafa; Nedosekin, Dmitry A.; Zharov, Vladimir P.

    2016-03-01

    In vivo flow cytometry using vessels as natural tubes with native cell flows has revolutionized the study of rare circulating tumor cells in a complex blood background. However, the presence of many blood cells in the detection volume makes it difficult to count each cell in this volume. We introduce method for manipulation of circulating cells in vivo with the use of gradient acoustic forces induced by ultrasound and photoacoustic waves. In a murine model, we demonstrated cell trapping, redirecting and focusing in blood and lymph flow into a tight stream, noninvasive wall-free transportation of blood, and the potential for photoacoustic detection of sickle cells without labeling and of leukocytes targeted by functionalized nanoparticles. Integration of cell focusing with intravital imaging methods may provide a versatile biological tool for single-cell analysis in circulation, with a focus on in vivo needleless blood tests, and preclinical studies of human diseases in animal models.

  7. Visually Relating Gene Expression and in vivo DNA Binding Data

    SciTech Connect

    Huang, Min-Yu; Mackey, Lester; Ker?,; nen, Soile V. E.; Weber, Gunther H.; Jordan, Michael I.; Knowles, David W.; Biggin, Mark D.; Hamann, Bernd

    2011-09-20

    Gene expression and in vivo DNA binding data provide important information for understanding gene regulatory networks: in vivo DNA binding data indicate genomic regions where transcription factors are bound, and expression data show the output resulting from this binding. Thus, there must be functional relationships between these two types of data. While visualization and data analysis tools exist for each data type alone, there is a lack of tools that can easily explore the relationship between them. We propose an approach that uses the average expression driven by multiple of ciscontrol regions to visually relate gene expression and in vivo DNA binding data. We demonstrate the utility of this tool with examples from the network controlling early Drosophila development. The results obtained support the idea that the level of occupancy of a transcription factor on DNA strongly determines the degree to which the factor regulates a target gene, and in some cases also controls whether the regulation is positive or negative.

  8. Tracking immune cells in vivo using magnetic resonance imaging

    PubMed Central

    Ahrens, Eric T.; Bulte, Jeff W. M.

    2013-01-01

    The increasing complexity of in vivo imaging technologies, coupled with the development of cell therapies, has fuelled a revolution in immune cell tracking in vivo. Powerful magnetic resonance imaging (MRI) methods are now being developed that use iron oxide- and 19F-based probes. These MRI technologies can be used for image-guided immune cell delivery and for the visualization of immune cell homing and engraftment, inflammation, cell physiology and gene expression. MRI-based cell tracking is now also being applied to evaluate therapeutics that modulate endogenous immune cell recruitment and to monitor emerging cellular immunotherapies. These recent uses show that MRI has the potential to be developed in many applications to follow the fate of immune cells in vivo. PMID:24013185

  9. In vivo acoustic and photoacoustic focusing of circulating cells

    PubMed Central

    Galanzha, Ekaterina I.; Viegas, Mark G.; Malinsky, Taras I.; Melerzanov, Alexander V.; Juratli, Mazen A.; Sarimollaoglu, Mustafa; Nedosekin, Dmitry A.; Zharov, Vladimir P.

    2016-01-01

    In vivo flow cytometry using vessels as natural tubes with native cell flows has revolutionized the study of rare circulating tumor cells in a complex blood background. However, the presence of many blood cells in the detection volume makes it difficult to count each cell in this volume. We introduce method for manipulation of circulating cells in vivo with the use of gradient acoustic forces induced by ultrasound and photoacoustic waves. In a murine model, we demonstrated cell trapping, redirecting and focusing in blood and lymph flow into a tight stream, noninvasive wall-free transportation of blood, and the potential for photoacoustic detection of sickle cells without labeling and of leukocytes targeted by functionalized nanoparticles. Integration of cell focusing with intravital imaging methods may provide a versatile biological tool for single-cell analysis in circulation, with a focus on in vivo needleless blood tests, and preclinical studies of human diseases in animal models. PMID:26979811

  10. Glucocorticoids enhance the in vivo migratory response of human monocytes.

    PubMed

    Yeager, Mark P; Pioli, Patricia A; Collins, Jane; Barr, Fiona; Metzler, Sara; Sites, Brian D; Guyre, Paul M

    2016-05-01

    Glucocorticoids (GCs) are best known for their potent anti-inflammatory effects. However, an emerging model for glucocorticoid (GC) regulation of in vivo inflammation also includes a delayed, preparatory effect that manifests as enhanced inflammation following exposure to an inflammatory stimulus. When GCs are transiently elevated in vivo following exposure to a stressful event, this model proposes that a subsequent period of increased inflammatory responsiveness is adaptive because it enhances resistance to a subsequent stressor. In the present study, we examined the migratory response of human monocytes/macrophages following transient in vivo exposure to stress-associated concentrations of cortisol. Participants were administered cortisol for 6h to elevate in vivo cortisol levels to approximate those observed during major systemic stress. Monocytes in peripheral blood and macrophages in sterile inflammatory tissue (skin blisters) were studied before and after exposure to cortisol or placebo. We found that exposure to cortisol induced transient upregulation of monocyte mRNA for CCR2, the receptor for monocyte chemotactic protein-1 (MCP-1/CCL2) as well as for the chemokine receptor CX3CR1. At the same time, mRNA for the transcription factor IκBα was decreased. Monocyte surface expression of CCR2 but not CX3CR1 increased in the first 24h after cortisol exposure. Transient exposure to cortisol also led to an increased number of macrophages and neutrophils in fluid derived from a sterile inflammatory site in vivo. These findings suggest that the delayed, pro-inflammatory effects of cortisol on the human inflammatory responses may include enhanced localization of effector cells at sites of in vivo inflammation. PMID:26790757

  11. In Vivo Gene Delivery by Nonviral Vectors: Overcoming Hurdles?

    PubMed Central

    Zhang, Yuan; Satterlee, Andrew; Huang, Leaf

    2012-01-01

    The promise of cancer gene therapeutics is hampered by difficulties in the in vivo delivery to the targeted tumor cells, and systemic delivery remains to be the biggest challenge to be overcome. Here, we concentrate on systemic in vivo gene delivery for cancer therapy using nonviral vectors. In this review, we summarize the existing delivery barriers together with the requirements and strategies to overcome these problems. We will also introduce the current progress in the design of nonviral vectors, and briefly discuss their safety issues. PMID:22525514

  12. Manipulating the in vivo immune response by targeted gene knockdown

    PubMed Central

    Lieberman, Judy

    2015-01-01

    Aptamers, nucleic acids selected for high affinity binding to proteins, can be used to activate or antagonize immune mediators or receptors in a location and cell-type specific manner and to enhance antigen presentation. They can also be linked to other molecules (other aptamers, siRNAs or miRNAs, proteins, toxins) to produce multifunctional compounds for targeted immune modulation in vivo. Aptamer-siRNA chimeras (AsiCs) that induce efficient cell-specific knockdown in immune cells in vitro and in vivo can be used as an immunological research tool or potentially as an immunomodulating therapeutic. PMID:26149459

  13. Manipulating the in vivo immune response by targeted gene knockdown.

    PubMed

    Lieberman, Judy

    2015-08-01

    Aptamers, nucleic acids selected for high affinity binding to proteins, can be used to activate or antagonize immune mediators or receptors in a location and cell-type specific manner and to enhance antigen presentation. They can also be linked to other molecules (other aptamers, siRNAs or miRNAs, proteins, toxins) to produce multifunctional compounds for targeted immune modulation in vivo. Aptamer-siRNA chimeras (AsiCs) that induce efficient cell-specific knockdown in immune cells in vitro and in vivo can be used as an immunological research tool or potentially as an immunomodulating therapeutic. PMID:26149459

  14. Toward quantitative "in vivo biochemistry" with fluorescence fluctuation spectroscopy.

    PubMed

    Slaughter, Brian D; Li, Rong

    2010-12-01

    Quantitative description of protein dynamics and interactions in vivo with temporal and spatial resolution is a key step in dissecting molecular mechanisms in cell biology. Fluorescence fluctuation spectroscopy (FFS) has recently emerged as a powerful in vivo tool for assessing molecular concentration and movement and formation of hetero- and homo-oligomeric complexes. This article discusses point FFS-based analysis methods that have proven useful to cell biologists, focusing on the kinds of information they provide, their pros and cons, and the basic instrumentation required. Along the way, we describe briefly a few recent examples where these analyses have helped address important biological questions. PMID:21160072

  15. Vaginal Lactobacillus: biofilm formation in vivo – clinical implications

    PubMed Central

    Ventolini, Gary

    2015-01-01

    Vaginal lactobacilli provide protection against intrusive pathogenic bacteria. Some Lactobacillus spp. produce in vitro a thick, protective biofilm. We report in vivo formation of biofilm by vaginal Lactobacillus jensenii. The biofilm formation was captured in fresh wet-mount microscopic samples from asymptomatic patients after treatment for recurrent bacterial vaginitis. In vivo documentation of biofilm formation is in our opinion noteworthy, and has significant clinical implications, among which are the possibility to isolate, grow, and therapeutically utilize lactobacilli to prevent recurrent vaginal infections and preterm labor associated with vaginal microbial pathogens. PMID:25733930

  16. Imaging targeted-agent binding in vivo with two probes

    NASA Astrophysics Data System (ADS)

    Pogue, Brian W.; Samkoe, Kimberley S.; Hextrum, Shannon; O'Hara, Julia A.; Jermyn, Michael; Srinivasan, Subhadra; Hasan, Tayyaba

    2010-05-01

    An approach to quantitatively image targeted-agent binding rate in vivo is demonstrated with dual-probe injection of both targeted and nontargeted fluorescent dyes. Images of a binding rate constant are created that reveal lower than expected uptake of epidermal growth factor in an orthotopic xenograft pancreas tumor (2.3×10-5 s-1), as compared to the normal pancreas (3.4×10-5 s-1). This approach allows noninvasive assessment of tumor receptor targeting in vivo to determine the expected contrast, spatial localization, and efficacy in therapeutic agent delivery.

  17. Application of in vivo laser scanning microscope in dermatology

    NASA Astrophysics Data System (ADS)

    Lademann, Juergen; Richter, H.; Otberg, N.; Lawrenz, F.; Blume-Peytavi, U.; Sterry, W.

    2003-10-01

    The state of the art of in-vivo and in-vitro penetration measurements of topically applied substances is described. Only optical techniques represent online measuring methods based on the absorption or scattering properties of the topically applied substances. Laser scanning microscopy (LSM) has become a promising method for investigations in dermatology and skin physiology, after it was possible to analyze the skin surface on any body side in-vivo. In the present paper the application of a dermatological laser scanning microscope for penetration and distribution measurements of topically applied substances is described. The intercellular and follicular penetration pathways were studied.

  18. In vivo tissue-wide synchronization of mitochondrial metabolic oscillations

    PubMed Central

    Porat-Shliom, Natalie; Chen, Yun; Tora, Muhibullah; Shitara, Akiko; Masedunskas, Andrius; Weigert, Roberto

    2014-01-01

    Summary Little is known about the spatio-temporal coordination of mitochondrial metabolism in multicellular organisms in situ. Using intravital microscopy in live animals, we here report that mitochondrial metabolism undergoes rapid and periodic oscillations under basal conditions. Notably, mitochondria in vivo behave as a network of functionally coupled oscillators, which maintain a high level of coordination throughout the tissue via the activity of gap junctions. These findings reveal a unique aspect of the relationship between tissue architecture and self-organization of mitochondrial metabolism in vivo. PMID:25373899

  19. In vivo melanoma depth detection by a handheld photoacoustic microscope

    NASA Astrophysics Data System (ADS)

    Zhou, Yong; Xing, Wenxin; Maslov, Konstantin I.; Cornelius, Lynn A.; Wang, Lihong V.

    2015-03-01

    We developed a handheld photoacoustic microscope (PAM) to detect melanoma and determine tumor depth in nude mice in vivo. Compared to our previous PAM system for melanoma imaging, a new light delivery mechanism is introduced to improve light penetration. We show that melanomas with 4.1 mm and 3.3 mm thicknesses can be successfully detected in phantom and in vivo experiments, respectively. With its deep melanoma imaging ability and novel handheld design, this system is promising for clinical melanoma diagnosis, prognosis, and surgical planning for patients at the bedside.

  20. OCT-based in vivo tissue injury mapping

    NASA Astrophysics Data System (ADS)

    Baran, Utku; Li, Yuandong; Wang, Ruikang K.

    2016-03-01

    Tissue injury mapping (TIM) is developed by using a non-invasive in vivo optical coherence tomography to generate optical attenuation coefficient and microvascular map of the injured tissue. Using TIM, the infarct region development in mouse cerebral cortex during stroke is visualized. Moreover, we demonstrate the in vivo human facial skin structure and microvasculature during an acne lesion development. The results indicate that TIM may help in the study and the treatment of various diseases by providing high resolution images of tissue structural and microvascular changes.

  1. In vivo activation and functions of the protease factor XII.

    PubMed

    Björkqvist, Jenny; Nickel, Katrin F; Stavrou, Evi; Renné, Thomas

    2014-11-01

    Combinations of proinflammatory and procoagulant reactions are the unifying principle for a variety of disorders affecting the cardiovascular system. Factor XII (FXII, Hageman factor) is a plasma protease that initiates the contact system. The biochemistry of the contact system in vitro is well understood; however, its in vivo functions are just beginning to emerge. The current review concentrates on activators and functions of the FXII-driven contact system in vivo. Elucidating its physiologic activities offers the exciting opportunity to develop strategies for the safe interference with both thrombotic and inflammatory diseases. PMID:25187064

  2. Biodegradable Xylitol-Based Elastomers: In Vivo Behavior and Biocompatibility

    PubMed Central

    Bruggeman, Joost P.; Bettinger, Christopher J.; Langer, Robert

    2010-01-01

    Biodegradable elastomers based on polycondensation reactions of xylitol with sebacic acid, referred to as poly(xylitol sebacate) (PXS) elastomers have recently been developed. Herein, we describe the in vivo behavior of PXS elastomers. Four PXS elastomers were synthesized, characterized and compared to poly(L-lactic-co-glycolic acid) (PLGA). PXS elastomers displayed a high level of structural integrity and form stability during degradation. The in vivo half-life ranged from approximately 3 to 52 weeks. PXS elastomers exhibited increased biocompatibility compared to PLGA implants. PMID:20540093

  3. Detecting and monitoring NO, SNO and nitrite in vivo

    PubMed Central

    Bellavia, Landon; Kim-Shapiro, Daniel B; King, S Bruce

    2015-01-01

    The detection and quantification of nitric oxide and related reactive nitrogen species in vivo is vital to the understanding of the pathology and/or treatment of numerous conditions. To that end, several detection and quantification methods have been developed to study NO, as well as its redox relatives, nitrite and S-nitrosothiols. While no single technique can offer a complete picture of the nitrogen cycle in a given system in vivo, familiarity with the benefits and limitations of several common tools for NOx determination can assist in the development of new diagnostics and therapeutics. PMID:26848400

  4. Opto-ultrasound imaging in vivo in deep tissue

    NASA Astrophysics Data System (ADS)

    Si, Ke; YanXu; Zheng, Yao; Zhu, Xinpei; Gong, Wei

    2016-02-01

    It is of keen importance of deep tissue imaging with high resolution in vivo. Here we present an opto-ultrasound imaging method which utilizes an ultrasound to confine the laser pulse in a very tiny spot as a guide star. The results show that the imaging depth is 2mm with a resolution of 10um. Meanwhile, the excitation power we used is less than 2mW, which indicates that our methods can be applied in vivo without optical toxicity and optical bleaching due to the excitation power.

  5. Tracking of Multimodal Therapeutic Nanocomplexes Targeting Breast Cancer in Vivo

    PubMed Central

    Bardhan, Rizia; Chen, Wenxue; Bartels, Marc; Perez-Torres, Carlos; Botero, Maria F.; McAninch, Robin Ward; Contreras, Alejandro; Schiff, Rachel; Pautler, Robia G.; Halas, Naomi J.; Joshi, Amit

    2014-01-01

    Nanoparticle-based therapeutics with local delivery and external electromagnetic field modulation holds extraordinary promise for soft-tissue cancers such as breast cancer; however, knowledge of the distribution and fate of nanoparticles in vivo is crucial for clinical translation. Here we demonstrate that multiple diagnostic capabilities can be introduced in photothermal therapeutic nanocomplexes by simultaneously enhancing both near-infrared fluorescence and magnetic resonance imaging (MRI). We track nanocomplexes in vivo, examining the influence of HER2 antibody targeting on nanocomplex distribution over 72 h. This approach provides valuable, detailed information regarding the distribution and fate of complex nanoparticles designed for specific diagnostic and therapeutic functions. PMID:21090693

  6. Recombination between linear double-stranded DNA substrates in vivo

    PubMed Central

    Narayanan, Kumaran; Sim, Edmund Ui-Hang; Ravin, Nikolai V.; Lee, Choon-Weng

    2009-01-01

    Recombineering technology in E. coli enables targeting of linear donor DNA to circular recipient DNA using short shared homology sequences. In this work, we demonstrate that recombineering is also able to support recombination between a pair of linear DNA substrates (linear/linear recombineering) in vivo in E. coli. Linear DNA up to 100 kb is accurately modified and remains intact without undergoing rearrangements after recombination. This system will be valuable for direct in vivo manipulation of large linear DNA including the N15 and PY54 prophages and linear animal viruses, and for assembly of linear constructs as artificial chromosome vectors. PMID:19454252

  7. In vivo Carbon-13 Nuclear Magnetic Resonance Studies of Mammals

    NASA Astrophysics Data System (ADS)

    Alger, J. R.; Sillerud, L. O.; Behar, K. L.; Gillies, R. J.; Shulman, R. G.; Gordon, R. E.; Shaw, D.; Hanley, P. E.

    1981-11-01

    Natural abundance carbon-13 nuclear magnetic resonances (NMR) from human arm and rat tissues have been observed in vivo. These signals arise primarily from triglycerides in fatty tissue. Carbon-13 NMR was also used to follow, in a living rat, the conversion of C-1--labeled glucose, which was introduced into the stomach, to C-1--labeled liver glycogen. The carbon-13 sensitivity and resolution obtained shows that natural abundance carbon-13 NMR will be valuable in the study of disorders in fat metabolism, and that experiments with substrates labeled with carbon-13 can be used to study carbohydrate metabolism in vivo.

  8. Questions about the behaviour of bacterial pathogens in vivo.

    PubMed Central

    Smith, H

    2000-01-01

    Bacterial pathogens cause disease in man and animals. They have unique biological properties, which enable them to colonize mucous surfaces, penetrate them, grow in the environment of the host, inhibit or avoid host defences and damage the host. The bacterial products responsible for these five biological requirements are the determinants of pathogenicity (virulence determinants). Current knowledge comes from studies in vitro, but now interest is increasing in how bacteria behave and produce virulence determinants within the infected host. There are three aspects to elucidate: bacterial activities, the host factors that affect them and the metabolic interactions between the two. The first is relatively easy to accomplish and, recently, new methods for doing this have been devised. The second is not easy because of the complexity of the environment in vivo and its ever-changing face. Nevertheless, some information can be gained from the literature and by new methodology. The third aspect is very difficult to study effectively unless some events in vivo can be simulated in vitro. The objectives of the Discussion Meeting were to describe the new methods and to show how they, and conventional studies, are revealing the activities of bacterial pathogens in vivo. This paper sets the scene by raising some questions and suggesting, with examples, how they might be answered. Bacterial growth in vivo is the primary requirement for pathogenicity. Without growth, determinants of the other four requirements are not formed. Results from the new methods are underlining this point. The important questions are as follows. What is the pattern of a developing infection and the growth rates and population sizes of the bacteria at different stages? What nutrients are present in vivo and how do they change as infection progresses and relate to growth rates and population sizes? How are these nutrients metabolized and by what bacterial mechanisms? Which bacterial processes handle

  9. Recent advances in microscopic techniques for visualizing leukocytes in vivo

    PubMed Central

    Jain, Rohit; Tikoo, Shweta; Weninger, Wolfgang

    2016-01-01

    Leukocytes are inherently motile and interactive cells. Recent advances in intravital microscopy approaches have enabled a new vista of their behavior within intact tissues in real time. This brief review summarizes the developments enabling the tracking of immune responses in vivo. PMID:27239292

  10. In vivo static field perturbations in magnetic resonance

    NASA Astrophysics Data System (ADS)

    Koch, Kevin Matthew

    2007-12-01

    Fundamental magnetic resonance (MR) theory assumes the spatial homogeneity of a dominating static magnetic field B = B 0ẑ. When this assumption is violated, a myriad of artifacts and compromising factors are introduced to MR spectra and images. Though in vivo nuclear magnetic resonance (NMR) is one of the most widely used scientific and diagnostic tools in medicine and biology, it remains haunted by the continual and persistant ghost of B0 inhomogeneity. An inclusive list of in vivo NMR applications severely impacted by B0 inhomogeneity could go on ad infinitum. Examples of such applications include neurosurgical utility in functional magnetic resonance imaging (fMRI), cerebral metabolic flux mapping, cerebral diffusion tractography, and abdominal diagnostic imaging. Given this wide impact on in vivo NMR, significant effort has been exerted in developing methods of compensating B0 inhomogeneity. Complicating this task is the sample-specific nature of in vivo B 0 inhomogeneity and its exacerbation with ever increasing B 0 field strengths. State of the art B 0 inhomogeneity compensation is currently at a critical juncture where homogenization demands are overwhelming the outer capabilities of existing technology and methods. This thesis addresses the B 0 inhomogeneity problem in the mammalian brain and presents novel solutions to the homogenization technology stalemate.

  11. Assessment of Glial Function in the In Vivo Retina

    PubMed Central

    Srienc, Anja I.; Kornfield, Tess E.; Mishra, Anusha; Burian, Michael A.; Newman, Eric A.

    2013-01-01

    Glial cells, traditionally viewed as passive elements in the CNS, are now known to have many essential functions. Many of these functions have been revealed by work on retinal glial cells. This work has been conducted almost exclusively on ex vivo preparations and it is essential that retinal glial cell functions be characterized in vivo as well. To this end, we describe an in vivo rat preparation to assess the functions of retinal glial cells. The retina of anesthetized, paralyzed rats is viewed with confocal microscopy and laser speckle flowmetry to monitor glial cell responses and retinal blood flow. Retinal glial cells are labeled with the Ca2+ indicator dye Oregon Green 488 BAPTA-1 and the caged Ca2+ compound NP-EGTA by injection of the compounds into the vitreous humor. Glial cells are stimulated by photolysis of caged Ca2+ and the activation state of the cells assessed by monitoring Ca2+ indicator dye fluorescence. We find that, as in the ex vivo retina, retinal glial cells in vivo generate both spontaneous and evoked intercellular Ca2+ waves. We also find that stimulation of glial cells leads to the dilation of neighboring retinal arterioles, supporting the hypothesis that glial cells regulate blood flow in the retina. This in vivo preparation holds great promise for assessing glial cell function in the healthy and pathological retina. PMID:22144328

  12. Veni, vidi, vici: in vivo molecular imaging of immune response.

    PubMed

    Gross, Shimon; Moss, Britney L; Piwnica-Worms, David

    2007-10-01

    "I came, I saw, I conquered," Julius Caesar proclaimed, highlighting the importance of direct visualization as a winning strategy. Continuing the "From the Field" series (see Editorial [2007] 26, 131), Gross et al. summarize how modern molecular imaging techniques can successfully dissect the complexities of immune response in vivo. PMID:17967405

  13. Strength in Numbers: Visualizing CTL-Mediated Killing In Vivo.

    PubMed

    Nolz, Jeffrey C; Hill, Ann B

    2016-02-16

    Cytotoxic CD8+ T lymphocytes (CTLs) have long been believed to be extremely efficient killers. Forster and colleagues (Halle et al., 2016) used in vivo imaging to tell a different story, in which each CTL killed only 2-16 targets a day, and several CTLs per target were needed to get the job done. PMID:26885849

  14. Educating and training India's next generation of in vivo pharmacologists

    PubMed Central

    Lewis, David I.

    2016-01-01

    The Indian Pharmaceutical Industry is undergoing rapid development and expansion. Critical to this process, and the future of drug discovery in India is the continued education and training of integrative or in vivo pharmacologists, equipped with the knowledge, skills and expertise to undertake studies using laboratory animals. Modern in vivo pharmacologists not only require manual or technical skills, but a much broader education including in animal welfare, ethics, the principles of the replacement, refinement and reduction of animals in research, and nonanimal alternative techniques. This education, training, and continued professional development throughout their careers can be provided, in part, through the use of online e-learning resources. While many excellent resources exist, they are hard to locate and not widely known to the community. To address this issue, Education and Training Resources in In vivo Sciences, a free website which provides access to free open access e-learning resources in in vivo pharmacology was developed. Use of this resource by researchers and educators will result in better-trained researchers and members of ethical review committees, which in turn will raise animal welfare standards, minimize the pain, suffering and distress of laboratory animals, and enhance scientific research. PMID:27298489

  15. 40 CFR 79.64 - In vivo micronucleus assay.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... information on this test guideline, the following references should be consulted. (1) 40 CFR 798.5395, In Vivo... mammalian cells. As the erythroblast develops into an erythrocyte (red blood cell), its main nucleus is... micronuclei is facilitated in these cells because they lack a main nucleus. (b) Definitions. For the...

  16. 40 CFR 79.64 - In vivo micronucleus assay.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... information on this test guideline, the following references should be consulted. (1) 40 CFR 798.5395, In Vivo... mammalian cells. As the erythroblast develops into an erythrocyte (red blood cell), its main nucleus is... micronuclei is facilitated in these cells because they lack a main nucleus. (b) Definitions. For the...

  17. 40 CFR 79.64 - In vivo micronucleus assay.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... information on this test guideline, the following references should be consulted. (1) 40 CFR 798.5395, In Vivo... mammalian cells. As the erythroblast develops into an erythrocyte (red blood cell), its main nucleus is... micronuclei is facilitated in these cells because they lack a main nucleus. (b) Definitions. For the...

  18. 40 CFR 79.64 - In vivo micronucleus assay.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... information on this test guideline, the following references should be consulted. (1) 40 CFR 798.5395, In Vivo... mammalian cells. As the erythroblast develops into an erythrocyte (red blood cell), its main nucleus is... micronuclei is facilitated in these cells because they lack a main nucleus. (b) Definitions. For the...

  19. 40 CFR 79.64 - In vivo micronucleus assay.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... information on this test guideline, the following references should be consulted. (1) 40 CFR 798.5395, In Vivo... mammalian cells. As the erythroblast develops into an erythrocyte (red blood cell), its main nucleus is... micronuclei is facilitated in these cells because they lack a main nucleus. (b) Definitions. For the...

  20. In-Vivo Zinc Metabolism by Isotope Ratio Mass Spectrometry

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The purpose of this chapter is to highlight some of the methodological and technical issues surrounding the in vivo use of stable isotopes and to provide examples of how such studies have advanced our knowledge of human zinc metabolism. The advantages and disadvantages of the currently available in...

  1. Recent advances in microscopic techniques for visualizing leukocytes in vivo.

    PubMed

    Jain, Rohit; Tikoo, Shweta; Weninger, Wolfgang

    2016-01-01

    Leukocytes are inherently motile and interactive cells. Recent advances in intravital microscopy approaches have enabled a new vista of their behavior within intact tissues in real time. This brief review summarizes the developments enabling the tracking of immune responses in vivo. PMID:27239292

  2. In vivo dermal absorption of pyrethroid pesticides in the rat.

    EPA Science Inventory

    The potential for exposure to pyrethroid pesticides has risen recently because of their increased use. The objective of this study was to examine the in vivo dermal absorption of bifenthrin, deltamethrin and permethrin in the rat. Hair on the dorsal side of anesthetized adult m...

  3. In Vivo Axial Loading of the Mouse Tibia

    PubMed Central

    Melville, Katherine M.; Robling, Alexander G.

    2015-01-01

    Summary Non-invasive methods to apply controlled, cyclic loads to the living skeleton are used as an anabolic agent to stimulate new bone formation in adults and enhance bone mass accrual in growing animals. These methods are also invaluable for understanding bone signaling pathways. Our focus here is on a particular loading model: in vivo axial compression of the mouse tibia. An advantage of loading the tibia is that changes are present in both the cancellous envelope of the proximal tibia and the cortical bone of the tibial diaphysis. To load the tibia of the mouse axially in vivo, a cyclic compressive load is applied up to five times a week to a single tibia per mouse for a duration lasting from 1 day to 6 weeks. With the contralateral limb as an internal control, the anabolic response of the skeleton to mechanical stimuli can be studied in a pairwise experimental design. Here, we describe the key parameters that must be considered before beginning an in vivo mouse tibial loading experiment, including methods for in vivo strain gauging of the tibial midshaft, and then we describe general methods for loading the mouse tibia for an experiment lasting multiple days. PMID:25331046

  4. Analysis of the mutations inducedd by conazole fungicides in vivo

    EPA Science Inventory

    The mouse liver tumorigenic conazo1e fungicides triadimefon and propiconazo1e have previously been shown to be in vivo mouse liver mutagens in the Big Blue" transgenic mutation assay when administered in feed at tumorigenic doses, whereas the nontumorigenic conazo1e myc1obutani1 ...

  5. Performance testing of radiobioassay laboratories: In vivo measurements, Final Report

    SciTech Connect

    MacLellan, J.A.; Traub, R.J.; Olsen, P.C.

    1990-04-01

    A study of two rounds of in vivo laboratory performance testing was undertaken by Pacific Northwest Laboratory (PNL) to determine the appropriateness of the in vivo performance criteria of draft American National Standards Institute (ANSI) standard ANSI N13.3, Performance Criteria for Bioassay.'' The draft standard provides guidance to in vivo counting facilities regarding the sensitivity, precision, and accuracy of measurements for certain categories of commonly assayed radionuclides and critical regions of the body. This report concludes the testing program by presenting the results of the Round Two testing. Testing involved two types of measurements: chest counting for radionuclide detection in the lung, and whole body counting for detection of uniformly distributed material. Each type of measurement was further divided into radionuclide categories as defined in the draft standard. The appropriateness of the draft standard criteria by measuring a laboratory's ability to attain them were judged by the results of both round One and Round Two testing. The testing determined that performance criteria are set at attainable levels, and the majority of in vivo monitoring facilities passed the criteria when complete results were submitted. 18 refs., 18 figs., 15 tabs.

  6. Detecting virulence and drug resistance mycobacterial phenotypes in vivo

    PubMed Central

    Timmins, Graham

    2015-01-01

    Bacterial phenotypes are predominantly studied in culture because detection of their specific metabolic pathways in the host is challenging. Development of stable isotope breath tests allowing in situ phenotype analyses may endow diagnostics with new modalities based upon direct monitoring of in vivo microbial metabolism and host–pathogen phenotypic interactions. PMID:25800730

  7. ASSESSING TCDD WASTING SYNDROME IN AN IN VIVO OBESITY MODEL

    EPA Science Inventory

    TCDD is a by-product of incineration commonly found as a microcontaminant in the food supply. The TCDD wasting syndrome, characterized by prolonged weight loss, has been examined for decades. Much of this work has focused on high dose in vivo and in vitro studies....

  8. A Comparison of In Vitro and In Vivo Asexual Embryogenesis.

    PubMed

    Hand, Melanie L; de Vries, Sacco; Koltunow, Anna M G

    2016-01-01

    In plants, embryogenesis generally occurs through the sexual process of double fertilization, which involves a haploid sperm cell fusing with a haploid egg cell to ultimately give rise to a diploid embryo. Embryogenesis can also occur asexually in the absence of fertilization, both in vitro and in vivo. Somatic or gametic cells are able to differentiate into embryos in vitro following the application of plant growth regulators or stress treatments. Asexual embryogenesis also occurs naturally in some plant species in vivo, from either ovule cells as part of a process defined as apomixis, or from somatic leaf tissue in other species. In both in vitro and in vivo asexual embryogenesis, the embryo precursor cells must attain an embryogenic fate without the act of fertilization. This review compares the processes of in vitro and in vivo asexual embryogenesis including what is known regarding the genetic and epigenetic regulation of each process, and considers how the precursor cells are able to change fate and adopt an embryogenic pathway. PMID:26619856

  9. Efficient in vivo Vascularization of Tissue Engineering Scaffolds

    PubMed Central

    Hegen, Anja; Blois, Anna; Tiron, Crina E.; Hellesøy, Monica; Micklem, David R.; Nör, Jacques E.; Akslen, Lars A.; Lorens, James B.

    2010-01-01

    The success of tissue engineering depends on the rapid and efficient formation of a functional blood vasculature. Adult blood vessels comprise endothelial cells and peri-vascular mural cells that assemble into patent tubules ensheathed by a basement membrane during angiogenesis. Using individual vessel components, we characterized intra-scaffold microvessel self-assembly efficiency in a physiological in vivo tissue engineering implant context. Primary human microvascular endothelial- and vascular smooth muscle cells were seeded at different ratios in poly-L lactic acid (PLLA) scaffolds enriched with basement membrane proteins (Matrigel) and implanted subcutaneously into immunocompromised mice. Temporal intra-scaffold microvessel formation, anastomosis and perfusion were monitored by immunohistochemical, flow cytometric and in vivo multiphoton fluorescence microscopy analysis. Vascularization in the tissue engineering context was strongly enhanced in the implants seeded with a complete complement of blood vessel components: Human microvascular endothelial and vascular smooth muscle cells in vivo assembled a patent microvasculature within Matrigel-enriched PLLA scaffolds that anastomosed with the host circulation during the first week of implantation. Multiphoton fluorescence angiographic analysis of the intra-scaffold microcirculation showed a uniform, branched microvascular network. 3-D image reconstruction analysis of hPASMC distribution within vascularized implants was non-random and displayed a preferential peri-vascular localization. Hence, efficient microvessel self-assembly, anastomosis and establishment of a functional microvasculture in the native hypoxic in vivo tissue engineering context is promoted by providing a complete set of vascular components. PMID:20865694

  10. Children's Coping with "In Vivo" Peer Rejection: An Experimental Investigation

    ERIC Educational Resources Information Center

    Reijntjes, Albert; Stegge, Hedy; Terwogt, Mark Meerum; Kamphuis, Jan Henk; Telch, Michael J.

    2006-01-01

    We examined children's behavioral coping in response to an "in vivo" peer rejection manipulation. Participants (N = 186) ranging between 10 and 13 years of age, played a computer game based on the television show "Survivor" and were randomized to either peer rejection (i.e., being voted out of the game) or non-rejection control. During a five-min.…

  11. ANTIOXIDANTS AMELIORATION OF ARSENICAL-INDUCED EFFECTS IN VIVO

    EPA Science Inventory

    Antioxidant amelioration of arsenical-induced effects in vivo. ES Hunter and EH Rogers. Reproductive Toxicology Division, NHEERL, US EPA, RTP, NC.

    Antioxidants have been reported to ameliorate the effects of many developmental toxicants. We tested the hypothesis that oxi...

  12. In vivo labeling of amyloid with BF-108.

    PubMed

    Suemoto, Takahiro; Okamura, Nobuyuki; Shiomitsu, Tsuyoshi; Suzuki, Masako; Shimadzu, Hiroshi; Akatsu, Hiroyasu; Yamamoto, Takayuki; Kudo, Yukitsuka; Sawada, Tohru

    2004-01-01

    Detection of aggregated amyloid-beta (Abeta) with a non-invasive imaging modality such as positron emission tomography (PET) was suggested to be ideal for the diagnosis of Alzheimer's disease (AD) prior to the onset of clinical symptoms. We have been searching for imaging probe candidates with a high affinity for aggregated Abeta in vitro and in vivo and high lipophilicity, a characteristic that allows for the permeation of the blood-brain barrier (BBB). As analyzed by Thioflavin T (ThT) assay and octanol/water partition coefficient test (PC), 3-diethylamino-6-(2-fluoroethyl)ethylaminoacridine (BF-108) were found to have high affinity for Abeta aggregates in vitro and high lipophilicity. Intravenously administrated BF-108 labeled Abeta aggregates injected into the amygdala as observed under a fluorescence microscope, showing this compound's permeability of BBB and an ability to label Abeta in vivo. BF-108 also labeled neuritic senile plaques (SPs), neurofibrillary tangles, and amyloid-laden vessels in temporal and hippocampal sections from AD patients. Following intravenous administration of BF-108 to an APP23 transgenic (TG) mouse, in vivo labeling of endogenous plaques was seen in brain sections by fluorescence microscopy. These properties suggest the potential utility of BF-108 for in vivo imaging of AD pathology. PMID:14687882

  13. Liposomes for scintigraphic imaging: optimization of in vivo behavior.

    PubMed

    Boerman, O C; Oyen, W J; Corstens, F H; Storm, G

    1998-12-01

    Liposomes, microscopic lipid vesicles consisting of concentric phospholipid bilayers enclosing discrete aqueous spaces, have been investigated extensively as carries for drugs in attempts to achieve selective deposition and/or reduced toxicity. Liposomes radiolabeled with gamma emitters (67Ga, 111In and 99mTc) have been used for imaging purposes. Liposomes as formulated in the past, are rapidly taken up by cells of the mononuclear phagocyte system, primarily those located in liver and spleen. However, it has been shown during the last two decades that the in vivo behavior of liposomes can be modulated by modifying their formulation. The size and the lipid composition have a major influence on the blood clearance rate, hepatic uptake and splenic uptake of liposomes. The development of long circulating liposomes, in particular coating of the bilayer with polyethyleneglycol (PEG) resulted in liposomes that oppose recognition by the MPS, thus displaying even longer circulatory half-lives. By carefully adjusting the liposomal formulation, the in vivo characteristics of liposomes can be tailored such that they become suitable vehicles for imaging various pathological processes in vivo. Liposomes have been proposed for tumor imaging, for infection imaging and as blood pool markers. Here, the factors that determine the in vivo behavior of liposomes and the current status of liposome-based radiopharmaceuticals are reviewed. PMID:9973842

  14. IN VITRO AND IN VIVO MUTAGENICITY STUDIES OF ENVIRONMENTAL CHEMICALS

    EPA Science Inventory

    The objectives of this project were to evaluate the mutagenicity of various compounds, mostly pesticides, using microbial and mammalian cell in vitro techniques, as well as in vivo techniques in Drosophila and mice, and to further develop and refine these procedures for applicati...

  15. Non-invasive in vivo measurement of macular carotenoids

    NASA Technical Reports Server (NTRS)

    Lambert, James L. (Inventor); Borchert, Mark S. (Inventor)

    2009-01-01

    A non-invasive in vivo method for assessing macular carotenoids includes performing Optical Coherence Tomography (OCT) on a retina of a subject. A spatial representation of carotenoid levels in the macula based on data from the OCT of the retina can be generated.

  16. Carbon Nanomaterials Interfacing with Neurons: An In vivo Perspective

    PubMed Central

    Baldrighi, Michele; Trusel, Massimo; Tonini, Raffaella; Giordani, Silvia

    2016-01-01

    Developing new tools that outperform current state of the art technologies for imaging, drug delivery or electrical sensing in neuronal tissues is one of the great challenges in neurosciences. Investigations into the potential use of carbon nanomaterials for such applications started about two decades ago. Since then, numerous in vitro studies have examined interactions between these nanomaterials and neurons, either by evaluating their compatibility, as vectors for drug delivery, or for their potential use in electric activity sensing and manipulation. The results obtained indicate that carbon nanomaterials may be suitable for medical therapies. However, a relatively small number of in vivo studies have been carried out to date. In order to facilitate the transformation of carbon nanomaterial into practical neurobiomedical applications, it is essential to identify and highlight in the existing literature the strengths and weakness that different carbon nanomaterials have displayed when probed in vivo. Unfortunately the current literature is sometimes sparse and confusing. To offer a clearer picture of the in vivo studies on carbon nanomaterials in the central nervous system, we provide a systematic and critical review. Hereby we identify properties and behavior of carbon nanomaterials in vivo inside the neural tissues, and we examine key achievements and potentially problematic toxicological issues. PMID:27375413

  17. Widefield quantitative multiplex surface enhanced Raman scattering imaging in vivo.

    PubMed

    McVeigh, Patrick Z; Mallia, Rupananda J; Veilleux, Israel; Wilson, Brian C

    2013-04-01

    In recent years numerous studies have shown the potential advantages of molecular imaging in vitro and in vivo using contrast agents based on surface enhanced Raman scattering (SERS), however the low throughput of traditional point-scanned imaging methodologies have limited their use in biological imaging. In this work we demonstrate that direct widefield Raman imaging based on a tunable filter is capable of quantitative multiplex SERS imaging in vivo, and that this imaging is possible with acquisition times which are orders of magnitude lower than achievable with comparable point-scanned methodologies. The system, designed for small animal imaging, has a linear response from (0.01 to 100 pM), acquires typical in vivo images in <10 s, and with suitable SERS reporter molecules is capable of multiplex imaging without compensation for spectral overlap. To demonstrate the utility of widefield Raman imaging in biological applications, we show quantitative imaging of four simultaneous SERS reporter molecules in vivo with resulting probe quantification that is in excellent agreement with known quantities (R²>0.98). PMID:23591913

  18. Radiosensitization in vitro and in vivo by 3-nitrotriazoles

    SciTech Connect

    Shibamoto, Y.; Sakano, K.; Kimura, R.; Nishidai, T.; Nishimoto, S.; Ono, K.; Kagiya, T.; Abe, M.

    1986-07-01

    A series of 3-nitro-1,2,4-triazole derivatives bearing various types of side chain (R) at the N1-position (AK-2000 series) were synthesized and their radiosensitizing effect and toxicity in vitro and in vivo were investigated, in comparison with those of Misonidazole (MISO), SR-2508, and RSU-1069. Of the fifteen 3-nitrotriazoles tested, all had sensitizing effects in vitro on hypoxic V79 cells. Also, all but one had definite effects on solid EMT6/KU and SCCVII tumors in vivo. For many of the triazole compounds, the degree of radiosensitization in vitro and in vivo appeared identical. However, they were generally less efficient, both in vitro and in vivo, than the corresponding 2-nitroimidazoles, whereas their aerobic cytotoxicity and toxicity to mice (LD50/7) were comparable to those of the 2-nitroimidazoles. Considering the sensitizing effect and toxicity, AK-2123 (R = CH/sub 2/CONHC/sub 2/H/sub 4/OCH/sub 3/) may be as useful as MISO, but none of the triazoles have been proved to be superior to SR-2508.

  19. Educating and training India's next generation of in vivo pharmacologists.

    PubMed

    Lewis, David I

    2016-01-01

    The Indian Pharmaceutical Industry is undergoing rapid development and expansion. Critical to this process, and the future of drug discovery in India is the continued education and training of integrative or in vivo pharmacologists, equipped with the knowledge, skills and expertise to undertake studies using laboratory animals. Modern in vivo pharmacologists not only require manual or technical skills, but a much broader education including in animal welfare, ethics, the principles of the replacement, refinement and reduction of animals in research, and nonanimal alternative techniques. This education, training, and continued professional development throughout their careers can be provided, in part, through the use of online e-learning resources. While many excellent resources exist, they are hard to locate and not widely known to the community. To address this issue, Education and Training Resources in In vivo Sciences, a free website which provides access to free open access e-learning resources in in vivo pharmacology was developed. Use of this resource by researchers and educators will result in better-trained researchers and members of ethical review committees, which in turn will raise animal welfare standards, minimize the pain, suffering and distress of laboratory animals, and enhance scientific research. PMID:27298489

  20. IN VIVO DERMAL ABSORPTION OF PYRETHROID PESTICIDES IN THE RAT

    EPA Science Inventory

    The potential for exposure to pyrethroid pesticides has risen recently because of their increased agricultural and residential use. The objective of this study was to examine the in vivo dermal absorption of bifenthrin, deltamethrin and cis-permethrin in the rat. Hair on...

  1. Monitoring tumor metastasis by in vivo imaging and flow cytometer

    NASA Astrophysics Data System (ADS)

    Gu, Zhenqin; Guo, Jin; Liu, Guangda; Li, Yan; Chen, Yun; Chen, Tong; Wang, Chen; Wei, Xunbin

    2009-08-01

    Prostate cancer is the most common malignancy in American men and the second leading cause of deaths from cancer, after lung cancer. The tumor usually grows slowly and remains confined to the gland for many years. During this time, the tumor produces little or no symptoms or outward signs. As the cancer advances, however, it can metastasize throughout other areas of the body, such as the bones, lungs, and liver. Surgical resection, hormonal therapy, chemotherapy and radiation therapy are the foundation of current prostate cancer therapies. Treatments for prostate cause both short- and long-term side effects that may be difficult to accept. Molecular mechanisms of prostate cancer metastasis need to be understood better and new therapies must be developed to selectively target to unique characteristics of cancer cell growth and metastasis. We have developed the "in vivo microscopy" to study the mechanisms that govern prostate cancer cell spread through the microenvironment in vivo in real-time confocal nearinfrared fluorescence imaging. A recently developed "in vivo flow cytometer" and optical imaging are used to assess prostate cancer cell spreading and the circulation kinetics of prostate cancer cells. A real- time quantitative monitoring of circulating prostate cancer cells by the in vivo flow cytometer will be useful to assess the effectiveness of the potential therapeutic interventions.

  2. In vivo Biodistribution and Clearance Studies using Multimodal ORMOSIL Nanoparticles

    PubMed Central

    Kumar, Rajiv; Roy, Indrajit; Ohulchanskky, Tymish Y.; Vathy, Lisa A.; Bergey, Earl J.; Sajjad, Munawwar; Prasad, Paras N

    2010-01-01

    Successful translation of the use of nanoparticles from laboratories to clinics requires exhaustive and elaborate studies involving the biodistribution, clearance and biocompatibility of nanoparticles for in vivo biomedical applications. We report here the use of multimodal organically modified silica (ORMOSIL) nanoparticles for in vivo bioimaging, biodistribution, clearance and toxicity studies. We have synthesized ORMOSIL nanoparticles with diameters of 20-25 nm, conjugated with near infra-red (NIR) fluorophores and radiolabelled them with 124I, for optical and PET imaging in vivo. The biodistribution of the non targeted nanoparticles was studied in non-tumored nude mice by optical fluorescence imaging, as well by measuring the radioactivity from harvested organs. Biodistribution studies showed a greater accumulation of nanoparticles in liver, spleen and stomach than in kidney, heart and lungs. The clearance studies carried out over a period of 15 days indicated hepatobiliary excretion of the nanoparticles. Selected tissues were analyzed for any potential toxicity by histological analysis, which confirmed the absence of any adverse effect or any other abnormalities in the tissues. The results demonstrate that these multimodal nanoparticles have potentially ideal attributes for use as biocompatible probes for in vivo imaging. PMID:20088598

  3. The morpholino molecular beacon for specific RNA visualization in vivo.

    PubMed

    Chen, Jianbin; Wu, Jikui; Hong, Yunhan

    2016-02-21

    A non-invasive fluorescent probe, morpholino molecular beacon (MO-MB), was designed for RNA visualization in vivo. Featuring negligible toxicity, stability, and high target specificity in living embryos, MO-MB is superior to conventional probes and has the potential for specific RNA visualization in basic biological and clinical research. PMID:26810703

  4. On-chip immobilization of planarians for in vivo imaging

    PubMed Central

    Dexter, Joseph P.; Tamme, Mary B.; Lind, Christine H.; Collins, Eva-Maria S.

    2014-01-01

    Planarians are an important model organism for regeneration and stem cell research. A complete understanding of stem cell and regeneration dynamics in these animals requires time-lapse imaging in vivo, which has been difficult to achieve due to a lack of tissue-specific markers and the strong negative phototaxis of planarians. We have developed the Planarian Immobilization Chip (PIC) for rapid, stable immobilization of planarians for in vivo imaging without injury or biochemical alteration. The chip is easy and inexpensive to fabricate, and worms can be mounted for and removed after imaging within minutes. We show that the PIC enables significantly higher-stability immobilization than can be achieved with standard techniques, allowing for imaging of planarians at sub-cellular resolution in vivo using brightfield and fluorescence microscopy. We validate the performance of the PIC by performing time-lapse imaging of planarian wound closure and sequential imaging over days of head regeneration. We further show that the device can be used to immobilize Hydra, another photophobic regenerative model organism. The simple fabrication, low cost, ease of use, and enhanced specimen stability of the PIC should enable its broad application to in vivo studies of stem cell and regeneration dynamics in planarians and Hydra. PMID:25227263

  5. In vivo Endothelial Cell Infection by Anaplasma marginale

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Anaplasma marginale has recently been shown to infect endothelial cells in vitro but it remains unknown as to whether endothelial infection also occurs in vivo. In this report, we demonstrate through dual fluorescence microscopy that A. marginale, detected by the monoclonal antibody, ANAF16C1, co-lo...

  6. In-vivo morphologic and spectroscopic investigation of Psoriasis

    NASA Astrophysics Data System (ADS)

    Kapsokalyvas, Dimitrios; Cicchi, Riccardo; Bruscino, Nicola; Alfieri, Domenico; Massi, Daniela; Lotti, Torello; Pavone, Francesco S.

    2011-07-01

    Psoriasis is an autoimmune disease of the skin characterized by hyperkeratosis, hyperproliferation of the epidermis, inflammatory cell accumulation and increased dilatation of dermal papillary blood vessels. Cases of psoriasis were investigated in vivo with optical means in order to evaluate the potential of in vivo optical biopsy. A Polarization Multispectral Dermoscope was employed for the macroscopic observation. Features such as the 'dotted' blood vessels pattern was observed with high contrast. High resolution image sections of the epidermis and the dermis were produced with a custom made Multiphoton Microscope. Imaging extended from the surface of the lesion down to the papillary dermis, at a depth of 200 μm. In the epidermis, a characteristic morphology of the stratum corneum found only in Psoriasis was revealed. Additionally, the cytoplasmic area of the cells in the stratum spinosum layer was found to be smaller than normal. In the dermis the morphological features were more pronounced, where the elongated dermal papillae dominated the papillary layer. Their length exceeds 100μm, which is a far greater value compared to that of healthy skin. These in vivo observations are consistent with the ex vivo histopathological observations, supporting both the applicability and potentiality of multispectral dermoscopy and multiphoton microscopy in the field of in vivo optical investigation and biopsy of skin.

  7. Methods of in-vivo mouse lung micro-CT

    NASA Astrophysics Data System (ADS)

    Recheis, Wolfgang A.; Nixon, Earl; Thiesse, Jacqueline; McLennan, Geoffrey; Ross, Alan; Hoffman, Eric

    2005-04-01

    Micro-CT will have a profound influence on the accumulation of anatomical and physiological phenotypic changes in natural and transgenetic mouse models. Longitudinal studies will be greatly facilitated, allowing for a more complete and accurate description of events if in-vivo studies are accomplished. The purpose of the ongoing project is to establish a feasible and reproducible setup for in-vivo mouse lung micro-computed tomography (μCT). We seek to use in-vivo respiratory-gated μCT to follow mouse models of lung disease with subsequent recovery of the mouse. Methodologies for optimizing scanning parameters and gating for the in-vivo mouse lung are presented. A Scireq flexiVent ventilated the gas-anesthetized mice at 60 breaths/minute, 30 cm H20 PEEP, 30 ml/kg tidal volume and provided a respiratory signal to gate a Skyscan 1076 μCT. Physiologic monitoring allowed the control of vital functions and quality of anesthesia, e.g. via ECG monitoring. In contrary to longer exposure times with ex-vivo scans, scan times for in-vivo were reduced using 35μm pixel size, 158ms exposure time and 18μm pixel size, 316ms exposure time to reduce motion artifacts. Gating via spontaneous breathing was also tested. Optimal contrast resolution was achieved at 50kVp, 200μA, applying an aluminum filter (0.5mm). There were minimal non-cardiac related motion artifacts. Both 35μm and 1μm voxel size images were suitable for evaluation of the airway lumen and parenchymal density. Total scan times were 30 and 65 minutes respectively. The mice recovered following scanning protocols. In-vivo lung scanning with recovery of the mouse delivered reasonable image quality for longitudinal studies, e.g. mouse asthma models. After examining 10 mice, we conclude μCT is a feasible tool evaluating mouse models of lung pathology in longitudinal studies with increasing anatomic detail available for evaluation as one moves from in-vivo to ex-vivo studies. Further developments include automated

  8. Clinical applications of in vivo fluorescence confocal laser scanning microscopy

    NASA Astrophysics Data System (ADS)

    Oh, Chilhwan; Park, Sangyong; Kim, Junhyung; Ha, Seunghan; Park, Gyuman; Lee, Gunwoo; Lee, Onseok; Chun, Byungseon; Gweon, Daegab

    2008-02-01

    Living skin for basic and clinical research can be evaluated by Confocal Laser Scanning Microscope (CLSM) non-invasively. CLSM imaging system can achieve skin image its native state either "in vivo" or "fresh biopsy (ex vivo)" without fixation, sectioning and staining that is necessary for routine histology. This study examines the potential fluorescent CLSM with a various exogenous fluorescent contrast agent, to provide with more resolution images in skin. In addition, in vivo fluorescent CLSM researchers will be extended a range of potential clinical application. The prototype of our CLSM system has been developed by Prof. Gweon's group. The operating parameters are composed of some units, such as illuminated wavelength 488 nm, argon illumination power up to 20mW on the skin, objective lens, 0.9NA oil immersion, axial resolution 1.0μm, field of view 200μm x 100μm (lateral resolution , 0.3μm). In human volunteer, fluorescein sodium was administrated topically and intradermally. Animal studies were done in GFP transgenic mouse, IRC mouse and pig skin. For imaging of animal skin, fluorescein sodium, acridine orange, and curcumine were used for fluorescein contrast agent. We also used the GFP transgenic mouse for fluorescein CLSM imaging. In intact skin, absorption of fluorescein sodium by individual corneocyte and hair. Intradermal administrated the fluorescein sodium, distinct outline of keratinocyte cell border could be seen. Curcumin is a yellow food dye that has similar fluorescent properties to fluorescein sodium. Acridin Orange can be highlight nuclei in viable keratinocyte. In vivo CLSM of transgenic GFP mouse enable on in vivo, high resolution view of GFP expressing skin tissue. GFP signals are brightest in corneocyte, kertinocyte, hair and eccrine gland. In intact skin, absorption of fluorescein sodium by individual corneocyte and hair. Intradermal administrated the fluorescein sodium, distinct outline of keratinocyte cell border could be seen. In

  9. Imaging cellular dynamics in vivo with multicolor fluorescent proteins

    NASA Astrophysics Data System (ADS)

    Hoffman, Robert M.

    2005-04-01

    The new field of in vivo cell biology is being developed with multi-colored fluorescent proteins. With the use of fluorescent proteins, the behavior of individual cells can be visualized in the living animal. An example of the new cell biology is dual-color fluorescence imaging using red fluorescent protein (RFP)-expressing tumors transplanted in green fluorescent protein (GFP)-expressing transgenic mice. These models show with great clarity the details of the tumor-stroma cell-cell interaction especially tumor-induced angiogenesis, tumor-infiltrating lymphocytes, stromal fibroblasts and macrophages. Another example is the color-coding of cells with RFP or GFP such that both cell types and their interaction can be simultaneously visualized in vivo. Stem cells can also be visualized and tracked in vivo with fluorescent proteins. Mice, in which the regulatory elements of the stem-cell marker nestin drive GFP expression, can be used to visualize hair follicle stem cells including their ability to form hair follicles as well as blood vessels. Dual-color cells expressing GFP in the nucleus and RFP in the cytoplasm enable real-time visualization of nuclear-cytoplasm dynamics including cell cycle events and apoptosis. Dual-color cells also enable the in vivo imaging of cell and nuclear deformation as well as trafficking in capillaries in living animals. Multiple-color labeling of cells will enable multiple events to be simultaneously visualized in vivo including cell-cell interaction, gene expression, ion fluxes, protein and organelle trafficking, chromosome dynamics and numerous other processes currently still studied in vitro.

  10. In Vivo Gene Delivery with L-Tyrosine Polyphosphate Nanoparticles

    PubMed Central

    Ditto, Andrew J.; Reho, John J.; Shah, Kush N.; Smolen, Justin A.; Holda, James H.; Ramirez, Rolando J.; Yun, Yang H.

    2013-01-01

    The concept of gene therapy is promising; however, the perceived risks and side effects associated with this technology have severely dampened the researchers’ enthusiasm.1–3 Thus, the development of a non-viral gene vector without immunological effects and with high transfection efficiency is necessary. Currently, most non-viral vectors have failed to achieve the in vivo transfection efficiencies of viral vectors due their toxicity,4 rapid clearance,5, 6 and/or inappropriate release rates.7 Although our previous studies have successfully demonstrated the controlled-release of plasmid DNA (pDNA) polyplexes encapsulated into nanoparticles formulated with L-tyrosine polyphosphate (LTP-pDNA nanoparticles),8, 9 the in vivo transfection capabilities and immunogenicity of this delivery system has yet to be examined. Thus, we evaluate LTP-pDNA nanoparticles in an in vivo setting via injection into rodent uterine tissue. Our results demonstrate through X-gal staining and immunohistochemistry of uterine tissue that transfection has successfully occurred after a nine-day incubation. In contrast, the results for the control nanoparticles show similar results to shams. Furthermore, reverse transcriptase polymerase chain reaction (RT-PCR) from the injected tissues confirms the transfection in vivo. To examine the immunogenicity, the LTP nanoparticles have been evaluated in a mouse model. No significant differences in the activation of the innate immune system are observed. These data provide the first report for the potential use of controlled-release nanoparticles formulated from an amino acid based polymer as an in vivo non-viral vector for gene therapy. PMID:23510151

  11. Dynamic conformal arc therapy: transmitted signal in vivo dosimetry.

    PubMed

    Piermattei, Angelo; Stimato, Gerardina; Gaudino, Diego; Ramella, Sara; D'Angelillo, Rolando Maria; Cellini, Francesco; Trodella, Lucio; D'Onofrio, Guido; Grimaldi, Luca; Cilla, Savino; Fidanzio, Andrea; Placidi, Elisa; Azario, Luigi

    2008-05-01

    A method for the determination of the in vivo isocenter dose, D(iso), has been applied to the dynamic conformal are therapy (DCAT) for thoracic tumors. The method makes use of the transmitted signal, S(t,alpha), measured at different gantry angles, a, by a small ion chamber positioned on the electronic portal imaging device. The in vivo method is implemented by a set of correlation functions obtained by the ratios between the transmitted signal and the midplane dose in a solid phantom, irradiated by static fields. The in vivo dosimetry at the isocenter for the DCAT requires the convolution between the signals, S(t,alpha), and the dose reconstruction factors, C(alpha), that depend on the patient's anatomy and on its tissue inhomogeneities along the beam central axis in the a direction. The C(alpha) factors are obtained by processing the patient's computed tomography scan. The method was tested by taking measurements in a cylindrical phantom and in a Rando Alderson phantom. The results show that the difference between the convolution calculations and the phantom measurements is within +/-2%. The in vivo dosimetry of the stereotactic DCAT for six lung tumors, irradiated with three or four arcs, is reported. The isocenter dose up to 17 Gy per therapy fraction was delivered on alternating days for three fractions. The agreement obtained in this pilot study between the total in vivo dose D(iso) and the planned dose D(iso,TPS) at the isocenter is +/-4%. The method has been applied on the DCAT obtaining a more extensive monitoring of possible systematic errors, the effect of which can invalidate the current therapy which uses a few high-dose fractions. PMID:18561658

  12. Dynamic conformal arc therapy: Transmitted signal in vivo dosimetry

    SciTech Connect

    Piermattei, Angelo; Stimato, Gerardina; Gaudino, Diego; Ramella, Sara; D'Angelillo, Rolando Maria; Cellini, Francesco; Trodella, Lucio; D'Onofrio, Guido; Grimaldi, Luca; Cilla, Savino; Fidanzio, Andrea; Placidi, Elisa; Azario, Luigi

    2008-05-15

    A method for the determination of the in vivo isocenter dose, D{sub iso}, has been applied to the dynamic conformal arc therapy (DCAT) for thoracic tumors. The method makes use of the transmitted signal, S{sub t,{alpha}}, measured at different gantry angles, {alpha}, by a small ion chamber positioned on the electronic portal imaging device. The in vivo method is implemented by a set of correlation functions obtained by the ratios between the transmitted signal and the midplane dose in a solid phantom, irradiated by static fields. The in vivo dosimetry at the isocenter for the DCAT requires the convolution between the signals , S{sub t,{alpha}}, and the dose reconstruction factors, C{sub {alpha}}, that depend on the patient's anatomy and on its tissue inhomogeneities along the beam central axis in the {alpha} direction. The C{sub {alpha}} factors are obtained by processing the patient's computed tomography scan. The method was tested by taking measurements in a cylindrical phantom and in a Rando Alderson phantom. The results show that the difference between the convolution calculations and the phantom measurements is within {+-}2%. The in vivo dosimetry of the stereotactic DCAT for six lung tumors, irradiated with three or four arcs, is reported. The isocenter dose up to 17 Gy per therapy fraction was delivered on alternating days for three fractions. The agreement obtained in this pilot study between the total in vivo dose D{sub iso} and the planned dose D{sub iso,TPS} at the isocenter is {+-}4%. The method has been applied on the DCAT obtaining a more extensive monitoring of possible systematic errors, the effect of which can invalidate the current therapy which uses a few high-dose fractions.

  13. Free-radical probes for functional in vivo EPR imaging

    NASA Astrophysics Data System (ADS)

    Subramanian, S.; Krishna, M. C.

    2007-02-01

    Electron paramagnetic resonance imaging (EPRI) is one of the recent functional imaging modalities that can provide valuable in vivo physiological information on its own merit and aids as a complimentary imaging technique to MRI and PET of tissues especially with respect to in vivo pO II (oxygen partial pressure), redox status and pharmacology. EPR imaging mainly deals with the measurement of distribution and in vivo dynamics and redox changes using special nontoxic paramagnetic spin probes that can be infused into the object of investigation. These spin probes should be characterized by simple EPR spectra, preferably with narrow EPR lines. The line width should be reversibly sensitive to the concentration of in vivo pO II with a linear dependence. Several non-toxic paramagnetic probes, some particulate and insoluble and others water-soluble and infusible (by intravenous or intramuscular injection) have been developed which can be effectively used to quantitatively assess tissue redox status, and tumor hypoxia. Quantitative assessment of the redox status of tissue in vivo is important in investigating oxidative stress, and that of tissue pO II is very important in radiation oncology. Other areas in which EPR imaging and oxymetry may help are in the investigation of tumorangiogenesis, wound healing, oxygenation of tumor tissue by the ingestion of oxygen-rich gases, etc. The correct choice of the spin probe will depend on the modality of measurement (whether by CW or time-domain EPR imaging) and the particular physiology interrogated. Examples of the available spin probes and some EPR imaging applications employing them are presented.

  14. Determination of Optimal Rhodamine Flurophore for In Vivo Optical Imaging

    PubMed Central

    Longmire, Michelle; Ogawa, Mikako; Hama, Yukihiro; Kosaka, Nobuyuki; Regino, Celeste A.S.; Choyke, Peter L.; Kobayashi, Hisataka

    2009-01-01

    Optical imaging has the potential to improve the efficacy of surgical and endoscopic approaches to cancer treatment; however, the optimal type of fluorescent probe has not yet been established. It is well known that rhodamine-core-derived fluorophores offer a combination of desirable properties such as good photostability, high extinction coefficient, and high fluorescence quantum yield. However, despite the ubiquitous use of rhodamine fluorophores for in vivo optical imaging, it remains to be determined, however, if unique chemical properties among individual rhodamine core family members affect fluorophore parameters critical to in vivo optical imaging applications. These parameters include: preserved fluorescence intensity in low pH environments, similar to that of the endolysosome; efficient fluorescence signal despite conformational changes to targeting proteins as may occur in harsh subcellular environments; persistence of fluorescence after cellular internalization; and sufficient signal-to-background ratios to permit the identification of fluorophore-targeted tumors. In the present study, we conjugated 4 common rhodamine-core based fluorescent dyes to a clinically feasible and quickly internalizing D-galactose receptor targeting reagent, galactosamine serum albumin (GmSA), and conducted a series of in vitro and in vivo experiments using a metastatic ovarian cancer mouse model to determine if differences exist among rhodamine fluorophores and if so, which rhodamine core possesses optimal characteristics for in vivo imaging applications. Herein, we demonstrate that the rhodamine-fluorophore, TAMRA, is the most robust of the 4 common rhodamine fluorophores for in vivo optical imaging of ovarian cancer metastases to the peritoneum. PMID:18610943

  15. Solvent casting flow from slot die

    NASA Astrophysics Data System (ADS)

    Lee, Semi; Nam, Jaewook

    2015-11-01

    A continuous solvent casting method using a slot die can precisely control the film thickness by adjusting the operating conditions, such as the belt speed and pumping rate, not the liquid property. Therefore, it is a suitable method for high precision continuous film production. In this particular method, the dope, or casting solution, is pumped through the feed slot to form a short curtain between the die and the moving belt. Although this method is widely used in producing films for various applications, it is difficult to find indepth analyses of such flow. In this study, we developed a finite element computational model for the steady-state two-dimensional sovent casting flow from the slot die. The effect of die configurations, rheological properties and operating conditions on the behavior and shape of the gas/liquid interfaces and the location of the dynamic contact line, which is the place where the dope meets the moving belt, were investigated.

  16. Things to Do After Someone Dies

    MedlinePlus

    ... are usually hidden by clothing. What about organ donation? At some time before death or right after ... lungs, pancreas, kidneys, cornea, liver, and skin. Organ donation allows healthy organs from someone who dies to ...

  17. Reinforced ceramic dies for superplastic forming operations

    NASA Astrophysics Data System (ADS)

    Sanders, Daniel G.

    2004-12-01

    Ceramic dies have been developed to meet the need for a dimensionally stable tool, which can withstand the temperatures (425 to 950 °C) and high forming pressures (up to 7 MPa) that are required for superplastic forming (SPF), superplastic forming with diffusion bonding (SPF/DB), and hot sizing of metal parts. With the improvements that have been made to strengthen fused silica based ceramics, the performance of ceramic tools is slowly closing in on meeting the same forming complexity as corrosion-resistant steel (CRES) dies can achieve. Boeing has successfully superplastically formed jet engine wide chord fan blades using ceramic dies, and many production aircraft parts are being built with Boeing’s patented ceramic die technology.

  18. Casting Characteristics of Aluminum Die Casting Alloys

    SciTech Connect

    Makhlouf M. Makhlouf; Diran Apelian

    2002-02-05

    The research program investigates the casting characteristics of selected aluminum die casting alloys. Specifically, the alloys' tendencies towards die soldering and sludge formation, and the alloys' fluidity and machinability are evaluated. It was found that: When the Fe and Mn contents of the alloy are low; caution has to be taken against possible die soldering. When the alloy has a high sludge factor, particularly a high level of Fe, measures must be taken to prevent the formation of large hardspots. For this kind of alloy, the Fe content should be kept at its lowest allowable level and the Mn content should be at its highest possible level. If there are problems in die filling, measures other than changing the alloy chemistry need to be considered first. In terms of alloy chemistry, the elements that form high temperature compounds must be kept at their lowest allowable levels. The alloys should not have machining problems when appropriate machining techniques and machining parameters are used.

  19. Geobasisdaten für die Planung?

    NASA Astrophysics Data System (ADS)

    Zölitz-Möller, Reinhard

    2002-09-01

    Die Nutzer von Geobasisdaten der Vermessungs- und Katasterverwaltungen finden heute vor allem in ATKIS (Amtliches Topographisch-Kartographisches Informationssystem) und in der ALK (automatisiert geführte Liegenschaftskarte; hier noch eingeschränkt) flächendeckende und für eine Fachdatenintegration geeignete Geodatenbestände vor. Gleichwohl wird von Nutzerseite häufig ein differenziertes und in Teilen kritisches Bild gezeichnet. Die Kritik richtet sich v.a. auf die Probleme, die ATKIS-Anwender mit dem Nutzerkomfort, dem komplexen Datenmodell, unrichtigen Flächennutzungsangaben, inkompatiblen Objektdefinitionen, den Preisen sowie mangelnder Aktualität und Vollständigkeit haben. Dennoch gibt es für viele Zwecke auch in der Planung langfristig keine echte Alternative zu den Geobasisdaten.

  20. Machining of Silicon-Ribbon-Forming Dies

    NASA Technical Reports Server (NTRS)

    Menna, A. A.

    1985-01-01

    Carbon extension for dies used in forming silicon ribbon crystals machined precisely with help of special tool. Die extension has edges beveled toward narrow flats at top, with slot precisely oriented and centered between flats and bevels. Cutting tool assembled from standard angle cutter and circular saw or saws. Angle cutters cuts bevels while slot saw cuts slot between them. In alternative version, custom-ground edges or additional circular saws also cut flats simultaneously.

  1. Thick film silicon growth techniques. [die materials

    NASA Technical Reports Server (NTRS)

    Bates, H. E.; Mlavsky, A. I.; Jewett, D. N.; White, V. E.

    1973-01-01

    The research which was directed toward finding an improved die material is reported. Wetting experiments were conducted with various materials to determine their compatibility with silicon. Work has also continued toward the development of quartz as a die material as new techniques have provided more optimistic results than observed in the past. As a result of the thermal modification previously described, improvements in growth stability have contributed to an increase in ribbon quality.

  2. Manufacture of die casting dies by hot isostatic pressing. CRADA final report

    SciTech Connect

    Viswanathan, S.; Ren, W.; Luk, K.; Brucher, H.G.

    1998-09-01

    The reason for this Cooperative Research and Development Agreement (CRADA) between the Oak Ridge National Laboratory (ORNL) and Doehler-Jarvis was to investigate the manufacture die-casting dies with internal water-cooling lines by hot-isostatic pressing (HIPing) of H13 tool steel powder. The use of HIPing will allow the near-net-shape manufacture of dies and the strategic placement of water-cooling lines during manufacture. The production of near-net-shape dies by HIPing involves the generation of HIPing diagrams, the design of the can that can be used for HIPing a die with complex details, strategic placement of water-cooling lines in the die, computer modeling to predict movement of the water lines during HIPing, and the development of strategies for placing water lines in the appropriate locations. The results presented include a literature review, particle analysis and characterization of H13 tool steel powder, and modeling of the HIPing process.

  3. Bromodeoxyuridine Inhibits Cancer Cell Proliferation In Vitro and In Vivo12

    PubMed Central

    Levkoff, Lindsay H; Marshall, Gregory P; Ross, Heather H; Caldeira, Maria; Reynolds, Brent A; Cakiroglu, Meryem; Mariani, Christopher L; Streit, Wolfgang J; Laywell, Eric D

    2008-01-01

    The thymidine analog bromodeoxyuridine (BrdU) is incorporated into newly synthesized DNA and has been shown to increase the susceptibility of incorporating cells to ionizing radiation. However, in the absence of secondary stressors, BrdU is thought to substitute relatively benignly for thymidine and is commonly used to “birth-date” proliferative cells. We report a novel antiproliferative effect of BrdU on cancer cells, which is independent of its role in radiosensitization. A single, brief in vitro exposure to BrdU induces a profound and sustained reduction in the proliferation rate of all cancer cells examined. Cells do not die but variably up-regulate some senescence-associated proteins as they accumulate in the G1 phase of the cell cycle. Bromodeoxyuridine also impairs the proliferative capacity of primary tumor-initiating human glioma cells and may therefore represent a means of targeting cancer stem cells. Finally, conservative in vivo BrdU regimens—in the absence of any other treatment—significantly suppress the progression of gliomas in the highly aggressive, syngeneic RG2 model. These results suggest that BrdU may have an important role as an adjunctive therapeutic for a wide variety of cancers based on new insights into its effect as a negative regulator of cell cycle progression. PMID:18680882

  4. Autophagy inhibition radiosensitizes in vitro, yet reduces radioresponses in vivo due to deficient immunogenic signalling.

    PubMed

    Ko, A; Kanehisa, A; Martins, I; Senovilla, L; Chargari, C; Dugue, D; Mariño, G; Kepp, O; Michaud, M; Perfettini, J-L; Kroemer, G; Deutsch, E

    2014-01-01

    Clinical oncology heavily relies on the use of radiotherapy, which often leads to merely transient responses that are followed by local or distant relapse. The molecular mechanisms explaining radioresistance are largely elusive. Here, we identified a dual role of autophagy in the response of cancer cells to ionizing radiation. On one hand, we observed that the depletion of essential autophagy-relevant gene products, such as ATG5 and Beclin 1, increased the sensitivity of human or mouse cancer cell lines to irradiation, both in vitro (where autophagy inhibition increased radiation-induced cell death and decreased clonogenic survival) and in vivo, after transplantation of the cell lines into immunodeficient mice (where autophagy inhibition potentiated the tumour growth-inhibitory effect of radiotherapy). On the other hand, when tumour proficient or deficient for autophagy were implanted in immunocompetent mice, it turned out that defective autophagy reduced the efficacy of radiotherapy. Indeed, radiotherapy elicited an anti-cancer immune response that was dependent on autophagy-induced ATP release from stressed or dying tumour cells and was characterized by dense lymphocyte infiltration of the tumour bed. Intratumoural injection of an ecto-ATPase inhibitor restored the immune infiltration of autophagy-deficient tumours post radiotherapy and improved the growth-inhibitory effect of ionizing irradiation. Altogether, our results reveal that beyond its cytoprotective function, autophagy confers immunogenic properties to tumours, hence amplifying the efficacy of radiotherapy in an immunocompetent context. This has far-reaching implications for the development of pharmacological radiosensitizers. PMID:24037090

  5. In vitro - in vivo correlation: from theory to applications.

    PubMed

    Emami, Jaber

    2006-01-01

    A key goal in pharmaceutical development of dosage forms is a good understanding of the in vitro and in vivo performance of the dosage forms. One of the challenges of biopharmaceutics research is correlating in vitro drug release information of various drug formulations to the in vivo drug profiles (IVIVC). Thus the need for a tool to reliably correlate in vitro and in vivo drug release data has exceedingly increased. Such a tool shortens the drug development period, economizes the resources and leads to improved product quality. Increased activity in developing IVIVCs indicates the value of IVIVCs to the pharmaceutical industry. IVIVC can be used in the development of new pharmaceuticals to reduce the number of human studies during the formulation development as the main objective of an IVIVC is to serve as a surrogate for in vivo bioavailability and to support biowaivers. It supports and/or validates the use of dissolution methods and specification settings. This is because the IVIVC includes in vivo relevance to in vitro dissolution specifications. It can also assist in quality control for certain scale-up and post-approval changes (SUPAC). With the proliferation of modified-release products, it becomes necessary to examine the concept of IVIVC in greater depth. Investigations of IVIVC are increasingly becoming an integral part of extended release drug development. There must be some in vitro means of assuring that each batch of the same product will perform identically in vivo. This review article represents the FDA guidance, development, evaluation, and validation of an IVIVC to grant biowaivers, and to set dissolution specifications for oral dosage forms, biopharmaceutics classification systems (BCS), BCS biowaivers, application of BCS in IVIVC development and concept of mapping. The importance of dissolution media and methodology and pharmacokinetic studies in the context of IVIVC has been highlighted. The review also covers the literature examples of IVIVCs

  6. Celestial Fireworks from Dying Stars

    NASA Astrophysics Data System (ADS)

    2011-04-01

    This image of the nebula NGC 3582, which was captured by the Wide Field Imager on the MPG/ESO 2.2-metre telescope at ESO's La Silla Observatory in Chile, shows giant loops of gas bearing a striking resemblance to solar prominences. These loops are thought to have been ejected by dying stars, but new stars are also being born within this stellar nursery. These energetic youngsters emit intense ultraviolet radiation that makes the gas in the nebula glow, producing the fiery display shown here. NGC 3582 is part of a large star-forming region in the Milky Way, called RCW 57. It lies close to the central plane of the Milky Way in the southern constellation of Carina (The Keel of Jason's ship, the Argo). John Herschel first saw this complex region of glowing gas and dark dust clouds in 1834, during his stay in South Africa. Some of the stars forming in regions like NGC 3582 are much heavier than the Sun. These monster stars emit energy at prodigious rates and have very short lives that end in explosions as supernovae. The material ejected from these dramatic events creates bubbles in the surrounding gas and dust. This is the probable cause of the loops visible in this picture. This image was taken through multiple filters. From the Wide Field Imager, data taken through a red filter are shown in green and red, and data taken through a filter that isolates the red glow characteristic of hydrogen are also shown in red. Additional infrared data from the Digitized Sky Survey are shown in blue. The image was processed by ESO using the observational data identified by Joe DePasquale, from the United States [1], who participated in ESO's Hidden Treasures 2010 astrophotography competition [2]. The competition was organised by ESO in October-November 2010, for everyone who enjoys making beautiful images of the night sky using astronomical data obtained using professional telescopes. Notes [1] Joe searched through ESO's archive and identified datasets that he used to compose his

  7. Do not let death do us part: 'find-me' signals in communication between dying cells and the phagocytes.

    PubMed

    Medina, C B; Ravichandran, K S

    2016-06-01

    The turnover and clearance of cells is an essential process that is part of many physiological and pathological processes. Improper or deficient clearance of apoptotic cells can lead to excessive inflammation and autoimmune disease. The steps involved in cell clearance include: migration of the phagocyte toward the proximity of the dying cells, specific recognition and internalization of the dying cell, and degradation of the corpse. The ability of phagocytes to recognize and react to dying cells to perform efficient and immunologically silent engulfment has been well-characterized in vitro and in vivo. However, how apoptotic cells themselves initiate the corpse removal and also influence the cells within the neighboring environment during clearance was less understood. Recent exciting observations suggest that apoptotic cells can attract phagocytes through the regulated release of 'find-me' signals. More recent studies also suggest that these find-me signals can have additional roles outside of phagocyte attraction to help orchestrate engulfment. This review will discuss our current understanding of the different find-me signals released by apoptotic cells, how they may be relevant in vivo, and their additional roles in facilitating engulfment. PMID:26891690

  8. 19F MRI for quantitative in vivo cell tracking

    PubMed Central

    Srinivas, Mangala; Heerschap, Arend; Ahrens, Eric T.; Figdor, Carl G.; de Vries, I. Jolanda M.

    2010-01-01

    Cellular therapy, including stem cell transplants and dendritic cell vaccines, is typically monitored for dosage optimization, accurate delivery and localization using non-invasive imaging, of which magnetic resonance imaging (MRI) is a key modality. 19F MRI retains the advantages of MRI as an imaging modality, while allowing direct detection of labelled cells for unambiguous identification and quantification, unlike typical metal-based contrast agents. Recent developments in 19F MRI-based in vivo cell quantification, the existing clinical use of 19F compounds and current explosive interest in cellular therapeutics have brought 19F imaging technology closer to clinical application. We review the application of 19F MRI to cell tracking, discussing intracellular 19F labels, cell labelling and in vivo quantification, as well as the potential clinical use of 19F MRI. PMID:20427096

  9. In vivo photoacoustic imaging of model of port wine stains.

    PubMed

    Yuan, Kaihua; Yuan, Yi; Gu, Ying; Gao, Jianhua; Xing, Da

    2012-01-01

    Port wine stains are categorized as a benign capillary vascular malformation, which is hard to cure. In this paper, a photoacoustic microscopy system, which integrated a two-dimensional scanning galvanometer, an objective lens and a focused ultrasound transducer, was designed for noninvasive imaging of blood vessels of port wine stains model in vivo. Cock comb was chosen as the port wine stains model in the experiment. The blood vessels in x-y plane and x-z plane were imaged clearly. Experimental results demonstrate that photoacoustic microscopy can image the blood vessels of port wine stains model in vivo with high contrast and high resolution. It has the potential for clinical applications in detecting the blood vessels in port wine stains skin. PMID:22635179

  10. In Vivo study of naturally deformed Escherichia coli bacteria.

    PubMed

    Tavaddod, Sharareh; Naderi-Manesh, Hossein

    2016-06-01

    A combination of light-microscopy and image processing has been applied to study naturally deformed Escherichia coli under in vivo condition and at the order of sub-pixel high-resolution accuracy. To classify deflagellated non-dividing E. coli cells to the rod-shape and bent-shape, a geometrical approach has been applied. From the analysis of the geometrical data which were obtained of image processing, we estimated the required effective energy for shaping a rod-shape to a bent-shape with the same size. We evaluated the energy of deformation in the naturally deformed bacteria with minimum cell manipulation, under in vivo condition, and with minimum influence of any external force, torque and pressure. Finally, we have also elaborated on the possible scenario to explain how naturally deformed bacteria are formed from initial to final-stage. PMID:27026097

  11. In vivo dynamics of RNA polymerase II transcription

    PubMed Central

    Darzacq, Xavier; Shav-Tal, Yaron; de Turris, Valeria; Brody, Yehuda; Shenoy, Shailesh M; Phair, Robert D; Singer, Robert H

    2016-01-01

    We imaged transcription in living cells using a locus-specific reporter system, which allowed precise, single-cell kinetic measurements of promoter binding, initiation and elongation. Photobleaching of fluorescent RNA polymerase II revealed several kinetically distinct populations of the enzyme interacting with a specific gene. Photobleaching and photoactivation of fluorescent MS2 proteins used to label nascent messenger RNAs provided sensitive elongation measurements. A mechanistic kinetic model that fits our data was validated using specific inhibitors. Polymerases elongated at 4.3 kilobases min−1, much faster than previously documented, and entered a paused state for unexpectedly long times. Transcription onset was inefficient, with only 1% of polymerase-gene interactions leading to completion of an mRNA. Our systems approach, quantifying both polymerase and mRNA kinetics on a defined DNA template in vivo with high temporal resolution, opens new avenues for studying regulation of transcriptional processes in vivo. PMID:17676063

  12. Nanoparticle PEBBLE sensors in live cells and in vivo

    PubMed Central

    Smith, Ron

    2009-01-01

    Nanoparticle sensors have been developed for imaging and dynamic monitoring, in live cells and in vivo, of the molecular or ionic components, constructs, forces and dynamics, all in real time, during biological/chemical/physical processes. With their biocompatible small size and inert matrix, nanoparticle sensors have been successfully applied for non-invasive real-time measurements of analytes and fields in cells and rodents, with spatial, temporal, physical and chemical resolution. This review describes the diverse designs of nanoparticle sensors for ions and small molecules, physical fields and biological features, as well as the characterization, properties, and applications of these nanosensors to in vitro and in vivo measurements. Their floating as well as localization ability in biological media is captured by the acronym PEBBLE: photonic explorer for bioanalysis with biologically localized embedding. PMID:20098636

  13. In-vivo absorption properties of algal pigments

    NASA Astrophysics Data System (ADS)

    Bidigare, Robert R.; Ondrusek, Michael E.; Morrow, John H.; Kiefer, Dale A.

    1990-09-01

    Estimates of the in vivo specific absorption coefficients (m2 mg'; 400-750 nm, 2 nm intervals) for the major algal pigment groups (chlorophylls, carotenoids and phycobilins) are presented. "Unpackaged" absorption coefficients were initially obtained by measuring the absorption properties of pure pigment standards spectrophotometrically and "shifting" their absorption maxima to match in vivo positions. Two approaches for estimating the phytoplankton absorption coefficient (spectral reconstruction and spectral decomposition) are compared by linear regression analysis, incorporating concurrent measurements of particulate absorption and pigmentation performed in the Sargasso Sea. Results suggest that "pigment package" effects are minimal for natural assemblages of open-oceanic phytoplankton and that accessory pigments do not always co-vary with chlorophyll a over depth and time.

  14. In Vivo Flow Cytometry: A Horizon of Opportunities

    PubMed Central

    Tuchin, Valery V.; Tárnok, Attila; Zharov, Vladimir P.

    2012-01-01

    Flow cytometry has been a fundamental tool of biological discovery for many years. Invasive extraction of cells from a living organism, however, may lead to changes in cell properties and prevents studying cells in their native environment. These problems can be overcome by use of in vivo flow cytometry which provides detection and imaging of circulating normal and abnormal cells directlyin blood or lymph flow. The goal of this mini-review is to provide a brief history, features and challenges of this new generation of flow cytometry methods and instruments. Spectrum of possibilities of in vivo flow cytometry in biological science (e.g., cell metabolism, immune function, or apoptosis) and medical fields (e.g., cancer, infection, cardiovascular disorder) including integrated photoacoustic-photothermal theranostics of circulating abnormal cells are discussed with focus on recent advances of this new platform. PMID:21915991

  15. Liposomes of terbutaline sulphate: in vitro and in vivo studies.

    PubMed

    Joshi, M R; Misra, A N

    1999-09-01

    In vitro studies were conducted to understand the comparative drug diffusion pattern, across artificial membrane, of the drug and of the prepared liposomes of different liposomal membrane composition. In vivo studies were carried out to determine the extent and time-course of pulmonary tissue uptake of administered liposomes containing terbutaline sulphate(TER) on rat lungs. In vitro studies revealed that the drug released from the prepared liposomes obeys Higuchi's diffusion controlled model. Different loading doses and release patterns of drug from the liposomes can be obtained by altering the PC:CHOL ratio and incorporation of cholesterol was found to reduce permeability of the membrane. Similarly drug absorption in vivo in rat's lung following intratracheal instillation, prolonged over 12 hr by liposomal entrapment of TER. The findings of present investigation indicated that liposomally encapsulated TER can be used for pulmonary delivery for maximizing the therapeutic efficacy and reducing undesirable side effects. PMID:10687283

  16. [Coeliac disease and reproduction: possible in vivo models].

    PubMed

    Stazi, Anna Velia

    2005-01-01

    Presently there are no in vivo models to study the different effects of coeliac disease (CD) including the increase of reproductive risks. CD is a multifactorial condition which requires both an exogenous element (gluten) and complex genetic factors; moreover, CD is associated to several endocrine, immune and reproductive diseases. There are no adequate in vivo models for the systemic complications of CD; in particular, there are no genetic knock-out models. However, models are available for gluten enteropathy such as Irish Setter and Balb/c and BDF1 mouse strains, and also for endocrine-immune diseases associated to CD such as BB rats and NOD mice. These models could be used to study reproductive aspects. This is desirable because a new model for dermatitis herpetiformis tightly associated with CD, that uses HLA-DQ8 transgenic NOD mice, has already been identified. PMID:16569922

  17. Label-free oxygen-metabolic photoacoustic microscopy in vivo

    PubMed Central

    Yao, Junjie; Maslov, Konstantin I.; Zhang, Yu; Xia, Younan; Wang, Lihong V.

    2011-01-01

    Almost all diseases, especially cancer and diabetes, manifest abnormal oxygen metabolism. Accurately measuring the metabolic rate of oxygen (MRO2) can be helpful for fundamental pathophysiological studies, and even early diagnosis and treatment of disease. Current techniques either lack high resolution or rely on exogenous contrast. Here, we propose label-free metabolic photoacoustic microscopy (mPAM) with small vessel resolution to noninvasively quantify MRO2in vivo in absolute units. mPAM is the unique modality for simultaneously imaging all five anatomical, chemical, and fluid-dynamic parameters required for such quantification: tissue volume, vessel cross-section, concentration of hemoglobin, oxygen saturation of hemoglobin, and blood flow speed. Hyperthermia, cryotherapy, melanoma, and glioblastoma were longitudinally imaged in vivo. Counterintuitively, increased MRO2 does not necessarily cause hypoxia or increase oxygen extraction. In fact, early-stage cancer was found to be hyperoxic despite hypermetabolism. PMID:21806264

  18. Digital image acquisition in in vivo confocal microscopy.

    PubMed

    Petroll, W M; Cavanagh, H D; Lemp, M A; Andrews, P M; Jester, J V

    1992-01-01

    A flexible system for the real-time acquisition of in vivo images has been developed. Images are generated using a tandem scanning confocal microscope interfaced to a low-light-level camera. The video signal from the camera is digitized and stored using a Gould image processing system with a real-time digital disk (RTDD). The RTDD can store up to 3200 512 x 512 pixel images at video rates (30 images s-1). Images can be input directly from the camera during the study, or off-line from a Super VHS video recorder. Once a segment of experimental interest is digitized onto the RTDD, the user can interactively step through the images, average stable sequences, and identify candidates for further processing and analysis. Examples of how this system can be used to study the physiology of various organ systems in vivo are presented. PMID:1552573

  19. Strategies to stabilize cell penetrating peptides for in vivo applications.

    PubMed

    Fominaya, Jesús; Bravo, Jerónimo; Rebollo, Angelita

    2015-10-01

    In the era of biomedicines and engineered carrier systems, cell penetrating peptides (CPPs) have been established as a promising tool for therapeutic application. Likewise, other therapeutic peptides, successful in vivo application of CPPs will strongly depend on peptide stability, the bottleneck for this type of biodegradable molecules. In this review, the authors describe the current knowledge of the in vivo degradation for known CPPs and the different strategies available to provide a higher resistance to metabolic degradation while preserving cell penetration efficiency. Peptide stability can be improved by different means, either modifying the structure to make it unrecognizable to proteases, or preventing access of proteolytic enzymes by applying conformation restriction or shielding strategies. PMID:26448473

  20. In vivo imaging of light-emitting probes

    NASA Astrophysics Data System (ADS)

    Rice, Bradley W.; Cable, Michael D.; Nelson, Michael B.

    2001-10-01

    In vivo imaging of cells tagged with light-emitting probes, such as firefly luciferase or fluorescent proteins, is a powerful technology that enables a wide range of biological studies in small research animals. Reporters with emission in the red to infrared (> 600 nm) are preferred due to the low absorption in tissue at these wavelengths. Modeling of photon diffusion through tissue indicates that bioluminescent cell counts as low as a few hundred can be detected subcutaneously, while approximately106 cells are required to detect signals at approximately 2 cm depth in tissue. Signal-to- noise estimates show that cooled back-thinned integrating charge coupled devices (CCDs) are preferred to image-intensified CCDs for this application, mainly due to their high quantum efficiency (approximately 85%) at wavelengths > 600 nm where tissue absorption is low. Instrumentation for in vivo imaging developed at Xenogen is described and several examples of images of mice with bioluminescent cells are presented.

  1. In Vivo Fluorescence Reflectance Imaging with Subcutaneous Mouse Tumor Models.

    PubMed

    Cao, Jie; Zhou, Mingzhou

    2016-01-01

    Optical imaging is undoubtedly one of the most versatile and widely used imaging techniques in both research and clinical practice. Among optical imaging technologies, fluorescence imaging is the most popularly used and has become an essential tool in biomedical science. A key component of fluorescence imaging is fluorescence-producing reporters, including fluorescent dyes and conjugates, as well as fluorescent proteins. For in vivo imaging applications, fluorophores with long emission at the near-infrared (NIR) region are generally preferred to overcome the photon attenuation in living tissue. Here, we describe the in vivo fluorescence imaging of an integrin αυβ3 targeted NIR fluorescent probe (cRGD-ICG-Der-02) using subcutaneous mouse tumor models. PMID:27283414

  2. New models for analyzing mast cell functions in vivo.

    PubMed

    Reber, Laurent L; Marichal, Thomas; Galli, Stephen J

    2012-12-01

    In addition to their well-accepted role as critical effector cells in anaphylaxis and other acute IgE-mediated allergic reactions, mast cells (MCs) have been implicated in a wide variety of processes that contribute to disease or help to maintain health. Although some of these roles were first suggested by analyses of MC products or functions in vitro, it is critical to determine whether, and under which circumstances, such potential roles actually can be performed by MCs in vivo. This review discusses recent advances in the development and analysis of mouse models to investigate the roles of MCs and MC-associated products during biological responses in vivo, and comments on some of the similarities and differences in the results obtained with these newer versus older models of MC deficiency. PMID:23127755

  3. A20-Deficient Mast Cells Exacerbate Inflammatory Responses In Vivo

    PubMed Central

    Vahl, J. Christoph; Aszodi, Attila; Peschke, Katrin; Schenten, Dominik; Hammad, Hamida; Beyaert, Rudi; Saur, Dieter; van Loo, Geert; Roers, Axel; Lambrecht, Bart N.; Kool, Mirjam; Schmidt-Supprian, Marc

    2014-01-01

    Mast cells are implicated in the pathogenesis of inflammatory and autoimmune diseases. However, this notion based on studies in mast cell-deficient mice is controversial. We therefore established an in vivo model for hyperactive mast cells by specifically ablating the NF-κB negative feedback regulator A20. While A20 deficiency did not affect mast cell degranulation, it resulted in amplified pro-inflammatory responses downstream of IgE/FcεRI, TLRs, IL-1R, and IL-33R. As a consequence house dust mite- and IL-33-driven lung inflammation, late phase cutaneous anaphylaxis, and collagen-induced arthritis were aggravated, in contrast to experimental autoimmune encephalomyelitis and immediate anaphylaxis. Our results provide in vivo evidence that hyperactive mast cells can exacerbate inflammatory disorders and define diseases that might benefit from therapeutic intervention with mast cell function. PMID:24453940

  4. In vivo behavior of a high performance duplex stainless steel.

    PubMed

    Cigada, A; De Santis, G; Gatti, A M; Roos, A; Zaffe, D

    1993-01-01

    An in vivo investigation of a new high molybdenum and nitrogen duplex stainless steel (25Cr--7Ni--4Mo--0.3N) has been performed. Cylindrical pins and specially developed devices, to test in static conditions the in vivo localized corrosion resistance, made of this new duplex steel and of a common austenitic stainless steel were implanted in rabbit's femurs for 6 and 12 months. After sacrifice, SEM observations and EDS microanalyses to detect metallic ion release were carried out on the femur sections surrounding the pins. Morphologic observations with stereoscope and SEM were performed on the metallic surfaces of the special devices in order to detect the presence of localized corrosion. Both ion release and localized corrosion were observed for the specimens made of austenitic stainless steel, but not for those made of 25Cr--7Ni--4Mo--0.3N duplex stainless steel. PMID:10148344

  5. In vivo recordings of brain activity using organic transistors

    PubMed Central

    Khodagholy, Dion; Doublet, Thomas; Quilichini, Pascale; Gurfinkel, Moshe; Leleux, Pierre; Ghestem, Antoine; Ismailova, Esma; Hervé, Thierry; Sanaur, Sébastien; Bernard, Christophe; Malliaras, George G.

    2013-01-01

    In vivo electrophysiological recordings of neuronal circuits are necessary for diagnostic purposes and for brain-machine interfaces. Organic electronic devices constitute a promising candidate because of their mechanical flexibility and biocompatibility. Here we demonstrate the engineering of an organic electrochemical transistor embedded in an ultrathin organic film designed to record electrophysiological signals on the surface of the brain. The device, tested in vivo on epileptiform discharges, displayed superior signal-to-noise ratio due to local amplification compared with surface electrodes. The organic transistor was able to record on the surface low-amplitude brain activities, which were poorly resolved with surface electrodes. This study introduces a new class of biocompatible, highly flexible devices for recording brain activity with superior signal-to-noise ratio that hold great promise for medical applications. PMID:23481383

  6. Applying the ARRIVE Guidelines to an In Vivo Database.

    PubMed

    Karp, Natasha A; Meehan, Terry F; Morgan, Hugh; Mason, Jeremy C; Blake, Andrew; Kurbatova, Natalja; Smedley, Damian; Jacobsen, Julius; Mott, Richard F; Iyer, Vivek; Matthews, Peter; Melvin, David G; Wells, Sara; Flenniken, Ann M; Masuya, Hiroshi; Wakana, Shigeharu; White, Jacqueline K; Lloyd, K C Kent; Reynolds, Corey L; Paylor, Richard; West, David B; Svenson, Karen L; Chesler, Elissa J; de Angelis, Martin Hrabě; Tocchini-Valentini, Glauco P; Sorg, Tania; Herault, Yann; Parkinson, Helen; Mallon, Ann-Marie; Brown, Steve D M

    2015-05-01

    The Animal Research: Reporting of In Vivo Experiments (ARRIVE) guidelines were developed to address the lack of reproducibility in biomedical animal studies and improve the communication of research findings. While intended to guide the preparation of peer-reviewed manuscripts, the principles of transparent reporting are also fundamental for in vivo databases. Here, we describe the benefits and challenges of applying the guidelines for the International Mouse Phenotyping Consortium (IMPC), whose goal is to produce and phenotype 20,000 knockout mouse strains in a reproducible manner across ten research centres. In addition to ensuring the transparency and reproducibility of the IMPC, the solutions to the challenges of applying the ARRIVE guidelines in the context of IMPC will provide a resource to help guide similar initiatives in the future. PMID:25992600

  7. In vitro and in vivo evaluation of SU-8 biocompatibility

    PubMed Central

    Nemani, Krishnamurthy V.; Moodie, Karen L.; Brennick, Jeoffry B.; Su, Alison; Gimi, Barjor

    2013-01-01

    SU-8 negative photoresist is a high tensile strength polymer that has been used for a number of biomedical applications that include cell encapsulation and neuronal probes. Chemically, SU-8 comprises, among other components, an epoxy based monomer and antimony salts, the latter being a potential source of cytotoxicity. We report on the in vitro and in vivo evaluation of SU-8 biocompatibility based on leachates from various solvents, at varying temperature and pH, and upon subcutaneous implantation of SU-8 substrates in mice. MTT cell viability assay did not exhibit any cytotoxic effects from the leachates. The hemolytic activity of SU-8 is comparable to that of FDA approved implant materials such as silicone elastomer, Buna-S and medical steel. In vivo histocompatibility study in mice indicates a muted immune response to subcutaneous SU-8 implants. PMID:23910365

  8. Verapamil inhibits scar formation after peripheral nerve repair in vivo

    PubMed Central

    Han, A-chao; Deng, Jing-xiu; Huang, Qi-shun; Zheng, Huai-yuan; Zhou, Pan; Liu, Zhi-wei; Chen, Zhen-bing

    2016-01-01

    The calcium channel blocker, verapamil, has been shown to reduce scar formation by inhibiting fibroblast adhesion and proliferation in vitro. It was not clear whether topical application of verapamil after surgical repair of the nerve in vivo could inhibit the formation of excessive scar tissue. In this study, the right sciatic nerve of adult Sprague-Dawley rats was transected and sutured with No. 10-0 suture. The stoma was wrapped with gelfoam soaked with verapamil solution for 4 weeks. Compared with the control group (stoma wrapped with gelfoam soaked with physiological saline), the verapamil application inhibited the secretion of extracellular matrix from fibroblasts in vivo, suppressed type I and III collagen secretion and increased the total number of axons and the number of myelinated axons. These findings suggest that verapamil could reduce the formation of scar tissue and promote axon growth after peripheral nerve repair. PMID:27127494

  9. Detection of Tight Junction Barrier Function In Vivo by Biotin

    PubMed Central

    Ding, Lei; Zhang, Yuguo; Tatum, Rodney; Chen, Yan-Hua

    2011-01-01

    Tight junctions (TJs) are the most apical component of the junctional complexes in mammalian epithelial cells and form selective paracellular barriers restricting the passage of solutes and ions across the epithelial sheets. Claudins, a TJ integral membrane protein family, play a critical role in regulating paracellular barrier permeability. In the in vitro cell culture system, transepithelial electrical resistance (TER) measurement and the flux of radioisotope or fluorescent labeled molecules with different sizes have been widely used to determine the TJ barrier function. In the in vivo system, the tracer molecule Sulfo-NHS-Biotin was initially used in Xenopus embryos system and subsequently was successfully applied to a number of animal tissues in situ and in different organisms under the experimental conditions to examine the functional integrity of TJs by several laboratories. In this chapter, we will describe the detailed procedures of applying biotin as a paracellular tracer molecule to different in vivo systems to assay TJ barrier function. PMID:21717351

  10. In vivo desensitization in the treatment of recurrent nightmares.

    PubMed

    Eccles, A; Wilde, A; Marshall, W L

    1988-12-01

    The literature, although sparse, suggests that behavioral interventions targeting the overt waking manifestations of the fear content of nightmares, can effectively reduce the frequency and intensity of these dreams. A 20-year-old female with recurrent nightmares centering on a fear of snakes, was treated by in vivo desensitization in which she approached a live, harmless snake. During treatment the client displayed habituation of fear of the snake and this corresponded to reductions in her nightmares. By the end of treatment and at 3-month and 1-year follow-up evaluations, she was free of fear and her nightmares had been eliminated. It is suggested that in vivo behavioral treatment of the content of nightmares may be a very effective way to eliminate these distressing dreams. PMID:2906945

  11. In vivo SELEX for Identification of Brain-penetrating Aptamers

    PubMed Central

    Cheng, Congsheng; Chen, Yong Hong; Lennox, Kim A; Behlke, Mark A; Davidson, Beverly L

    2013-01-01

    The physiological barriers of the brain impair drug delivery for treatment of many neurological disorders. One delivery approach that has not been investigated for their ability to penetrate the brain is RNA-based aptamers. These molecules can impart delivery to peripheral tissues and circulating immune cells, where they act as ligand mimics or can be modified to carry payloads. We developed a library of aptamers and an in vivo evolution protocol to determine whether specific aptamers could be identified that would home to the brain after injection into the peripheral vasculature. Unlike biopanning with recombinant bacteriophage libraries, we found that the aptamer library employed here required more than 15 rounds of in vivo selection for convergence to specific sequences. The aptamer species identified through this approach bound to brain capillary endothelia and penetrated into the parenchyma. The methods described may find general utility for targeting various payloads to the brain. PMID:23299833

  12. Molecular approaches for manipulating astrocytic signaling in vivo

    PubMed Central

    Xie, Alison X.; Petravicz, Jeremy; McCarthy, Ken D.

    2015-01-01

    Astrocytes are the predominant glial type in the central nervous system and play important roles in assisting neuronal function and network activity. Astrocytes exhibit complex signaling systems that are essential for their normal function and the homeostasis of the neural network. Altered signaling in astrocytes is closely associated with neurological and psychiatric diseases, suggesting tremendous therapeutic potential of these cells. To further understand astrocyte function in health and disease, it is important to study astrocytic signaling in vivo. In this review, we discuss molecular tools that enable the selective manipulation of astrocytic signaling, including the tools to selectively activate and inactivate astrocyte signaling in vivo. Lastly, we highlight a few tools in development that present strong potential for advancing our understanding of the role of astrocytes in physiology, behavior, and pathology. PMID:25941472

  13. In Vivo Assessment of Muscle Contractility in Animal Studies.

    PubMed

    Iyer, Shama R; Valencia, Ana P; Hernández-Ochoa, Erick O; Lovering, Richard M

    2016-01-01

    In patients with muscle injury or muscle disease, assessment of muscle damage is typically limited to clinical signs, such as tenderness, strength, range of motion, and more recently, imaging studies. Animal models provide unmitigated access to histological samples, which provide a "direct measure" of damage. However, even with unconstrained access to tissue morphology and biochemistry assays, the findings typically do not account for loss of muscle function. Thus, the most comprehensive measure of the overall health of the muscle is assessment of its primary function, which is to produce contractile force. The majority of animal models testing contractile force have been limited to the muscle groups moving the ankle, with advantages and disadvantages depending on the equipment. Here, we describe in vivo methods to measure torque, to produce a reliable muscle injury, and to follow muscle function within the same animal over time. We also describe in vivo methods to measure tension in the leg and thigh muscles. PMID:27492180

  14. In vivo red cell destruction by anti-Lu6

    SciTech Connect

    Issitt, P.D.; Valinsky, J.E.; Marsh, W.L.; DiNapoli, J.; Gutgsell, N.S. )

    1990-03-01

    An example is presented of an IgG1, anti-Lu6, that reacted by indirect antiglobulin test and was capable of destroying antigen-positive red cells in vivo. Two methods for the measurement of red cell survival, {sup 51}Cr labeling and flow cytometry, gave the same result: 20 percent of the test dose of Lu:6 red cells was destroyed in the first hour after injection and 80 percent in the first 24 hours. The clinical relevance of the antibody was correctly predicted by an in vitro monocyte monolayer assay. The finding that this example of anti-Lu6 was clinically significant should not be taken to mean that all antibodies directed against high-incidence Lutheran and Lutheran system-related antigens will behave similarly. When such antibodies are encountered, in vivo and/or in vitro studies to assess their clinical significance are necessary before rare blood is used for transfusion.

  15. Applying the ARRIVE Guidelines to an In Vivo Database

    PubMed Central

    Karp, Natasha A.; Meehan, Terry F.; Morgan, Hugh; Mason, Jeremy C.; Blake, Andrew; Kurbatova, Natalja; Smedley, Damian; Jacobsen, Julius; Mott, Richard F.; Iyer, Vivek; Matthews, Peter; Melvin, David G.; Wells, Sara; Flenniken, Ann M.; Masuya, Hiroshi; Wakana, Shigeharu; White, Jacqueline K.; Lloyd, K. C. Kent; Reynolds, Corey L.; Paylor, Richard; West, David B.; Svenson, Karen L.; Chesler, Elissa J.; de Angelis, Martin Hrabě; Tocchini-Valentini, Glauco P.; Sorg, Tania; Herault, Yann; Parkinson, Helen; Mallon, Ann-Marie; Brown, Steve D. M.

    2015-01-01

    The Animal Research: Reporting of In Vivo Experiments (ARRIVE) guidelines were developed to address the lack of reproducibility in biomedical animal studies and improve the communication of research findings. While intended to guide the preparation of peer-reviewed manuscripts, the principles of transparent reporting are also fundamental for in vivo databases. Here, we describe the benefits and challenges of applying the guidelines for the International Mouse Phenotyping Consortium (IMPC), whose goal is to produce and phenotype 20,000 knockout mouse strains in a reproducible manner across ten research centres. In addition to ensuring the transparency and reproducibility of the IMPC, the solutions to the challenges of applying the ARRIVE guidelines in the context of IMPC will provide a resource to help guide similar initiatives in the future. PMID:25992600

  16. Chemiluminescence detection from sonodynamic action in vitro and in vivo

    NASA Astrophysics Data System (ADS)

    He, Yonghong; Xing, Da; Yan, Guihong; Ueda, Ken-ichi

    2001-10-01

    In this work, chemiluminescence method was engaged for the first time to detect the active oxygen spices during sonocynamic action both in vitro and in vivo. We used CLA derivatives, which can efficiently react with singlet oxygen (1O2) or superoxide anion (O2-) to emit light, and luminol, which can be oxidized by a variety of free radicals to emit photons, to real-timely detect oxygen free radical formation in the sonosensitization of two sonosensitizer ATX-70 and HpD. The results show that 1O2 is involved in the sonosensitization. The mechanism of sonosensitizing was discussed. In vivo experiments, tumor imaging by sonodynamic chemiluminescence detection methods was established. This method could have potential applications in clinics for early-stage tumor diagnosis.

  17. Recent molecular approaches to understanding astrocyte function in vivo

    PubMed Central

    Davila, David; Thibault, Karine; Fiacco, Todd A.; Agulhon, Cendra

    2013-01-01

    Astrocytes are a predominant glial cell type in the nervous systems, and are becoming recognized as important mediators of normal brain function as well as neurodevelopmental, neurological, and neurodegenerative brain diseases. Although numerous potential mechanisms have been proposed to explain the role of astrocytes in the normal and diseased brain, research into the physiological relevance of these mechanisms in vivo is just beginning. In this review, we will summarize recent developments in innovative and powerful molecular approaches, including knockout mouse models, transgenic mouse models, and astrocyte-targeted gene transfer/expression, which have led to advances in understanding astrocyte biology in vivo that were heretofore inaccessible to experimentation. We will examine the recently improved understanding of the roles of astrocytes – with an emphasis on astrocyte signaling – in the context of both the healthy and diseased brain, discuss areas where the role of astrocytes remains debated, and suggest new research directions. PMID:24399932

  18. Infrared laser welding of the rabbit cornea in vivo

    NASA Astrophysics Data System (ADS)

    Williams, John M.; Burstein, Neal L.; Nowicki, Michael J.; Zietkiewicz, Christopher J.; Jeffers, William Q.

    1995-05-01

    The hydrogen fluoride laser has been used to successfully weld corneal tissue in vivo. Previous experiments have demonstrate the success of producing watertight welds in both porcine and human cadaver corneas. Wound bursting strengths of up to three times normal intraocular pressure have been reported. In this study, an in vivo model was utilized, specifically the rabbit cornea. Twelve New Zealand white rabbits, received a 7 mm, full thickness, linear corneal incision in one eye, and stay sutures were placed. Six of the wounds were welded with a semiconductor infrared laser, and six eyes served as controls. At two and four weeks, both histologic and tensiometric studies were performed. There was a trend toward increasing wound strength when the two and four week specimens were compared. Corneal welding may prove to be an adjunct to current suturing techniques in humans. Procedures requiring the closure of corneal incisions such as cataract extraction or penetrating keratoplasty may benefit from this technique.

  19. Non-contact intracellular binding of chloroplasts in vivo

    NASA Astrophysics Data System (ADS)

    Li, Yuchao; Xin, Hongbao; Liu, Xiaoshuai; Li, Baojun

    2015-06-01

    Non-contact intracellular binding and controllable manipulation of chloroplasts in vivo was demonstrated using an optical fiber probe. Launching a 980-nm laser beam into a fiber, which was placed about 3 μm above the surface of a living plant (Hydrilla verticillata) leaf, enabled stable binding of different numbers of chloroplasts, as well as their arrangement into one-dimensional chains and two-dimensional arrays inside the leaf without damaging the chloroplasts. Additionally, the formed chloroplast chains were controllably transported inside the living cells. The optical force exerted on the chloroplasts was calculated to explain the experimental results. This method provides a flexible method for studying intracellular organelle interaction with highly organized organelle-organelle contact in vivo in a non-contact manner.

  20. Three-Dimensional Cell Culture: A Breakthrough in Vivo

    PubMed Central

    Antoni, Delphine; Burckel, Hélène; Josset, Elodie; Noel, Georges

    2015-01-01

    Cell culture is an important tool for biological research. Two-dimensional cell culture has been used for some time now, but growing cells in flat layers on plastic surfaces does not accurately model the in vivo state. As compared to the two-dimensional case, the three-dimensional (3D) cell culture allows biological cells to grow or interact with their surroundings in all three dimensions thanks to an artificial environment. Cells grown in a 3D model have proven to be more physiologically relevant and showed improvements in several studies of biological mechanisms like: cell number monitoring, viability, morphology, proliferation, differentiation, response to stimuli, migration and invasion of tumor cells into surrounding tissues, angiogenesis stimulation and immune system evasion, drug metabolism, gene expression and protein synthesis, general cell function and in vivo relevance. 3D culture models succeed thanks to technological advances, including materials science, cell biology and bioreactor design. PMID:25768338

  1. In vivo photoacoustic imaging of osteosarcoma in a rat model

    NASA Astrophysics Data System (ADS)

    Hu, Jun; Yu, Menglei; Ye, Fei; Xing, Da

    2011-02-01

    Osteosarcoma is one of the most common primary malignant tumors of the bone and the second leading cause of cancer-related deaths in the pediatric age group. Confirmed diagnosis and prompt treatment of osteosarcoma are critical for effective prognosis. In this study, we investigate the application of photoacoustic imaging (PAI) for the detection of osteosarcoma in an animal model. Cross-section images of a normal rat leg and a tumorous rat leg were successfully reconstructed in vivo. Morphological changes and the development of the implanted osteosarcoma were accurately mapped with time-dependent photoacoustic images. Furthermore, we evaluate the use of gold nanorods as contrast agents for imaging osteosarcoma with PAI. This is the first study that uses PAI to detect osteosarcoma in vivo, and the results suggest that PAI has the potential clinical application for detecting osteosarcoma in the early stage.

  2. In vivo photoacoustic imaging of osteosarcoma on animal model

    NASA Astrophysics Data System (ADS)

    Yu, Menglei; Ye, Fei; Hu, Jun

    2011-01-01

    Osteosarcoma is the commonest primary malignant tumor of bone, and the second highest cause of cancer-related death in the paediatric age group. Although there are several methods for osteosarcoma detection, e.g. X-ray, CT, MRI and bone scan, they are not satisfied methods because they can hardly detect osteosarcoma in early stage. Photoacoustic imaging (PAI) is an emerging hybrid imaging modality that is noninvasive, nonionizing, with high sensitivity, satisfactory imaging depth and good temporal and spatial resolution. In order to explore this new method to detect osteosarcoma, we established SD rat models with osteosarcoma and utilized PAI to reconstruct the osteosarcoma image in vivo. This is the first time detecting osteosarcoma in vivo using PAI, and the results suggested that PAI has potential clinical application for detecting osteosarcoma in the early stage.

  3. Biotinylated erythrocytes: in vivo survival and in vitro recovery

    SciTech Connect

    Suzuki, T.; Dale, G.L.

    1987-09-01

    Rabbit erythrocytes were biotinylated by reaction with N-hydroxysuccinimidobiotin; the average level of biotinylation was 25,000 molecules per erythrocyte. These biotinylated cells exhibited a normal survival rate when reinfused into rabbits. Two studies demonstrated that the biotin label was stable in vivo. The first was a double-labeling experiment where the biotinylated erythrocytes were also labeled with /sup 14/C-cyanate; on reinfusion, the loss of biotinylated erythrocytes and /sup 14/C-cyanate label occurred in unison. The second study demonstrated that biotinylated erythrocytes that had been reinfused into rabbits could later be selectively isolated by attachment to an avidin support. This technique will facilitate a variety of studies that require the ability to label a specific cohort of cells in vitro and then reisolate those same cells after in vivo recirculation.

  4. Manipulating leukocyte interactions in vivo through optogenetic chemokine release.

    PubMed

    Sarris, Milka; Olekhnovitch, Romain; Bousso, Philippe

    2016-06-01

    Light-mediated release of signaling ligands, such as chemoattractants, growth factors, and cytokines is an attractive strategy for investigation and therapeutic targeting of leukocyte communication and immune responses. We introduce a versatile optogenetic method to control ligand secretion, combining UV-conditioned endoplasmic reticulum-to-Golgi trafficking and a furin-processing step. As proof of principle, we achieved light-triggered chemokine secretion and demonstrated that a brief pulse of chemokine release can mediate a rapid flux of leukocyte contacts with target cells in vitro and in vivo. This approach opens new possibilities for dynamic investigation of leukocyte communication in vivo and may confer the potential to control the local release of soluble mediators in the context of immune cell therapies. PMID:27057000

  5. In vivo excitation of nanoparticles using luminescent bacteria

    PubMed Central

    Dragavon, Joe; Blazquez, Samantha; Rekiki, Abdessalem; Samson, Chelsea; Theodorou, Ioanna; Rogers, Kelly L.; Tournebize, Régis; Shorte, Spencer L.

    2012-01-01

    The lux operon derived from Photorhabdus luminescens incorporated into bacterial genomes, elicits the production of biological chemiluminescence typically centered on 490 nm. The light-producing bacteria are widely used for in vivo bioluminescence imaging. However, in living samples, a common difficulty is the presence of blue-green absorbers such as hemoglobin. Here we report a characterization of fluorescence by unbound excitation from luminescence, a phenomenon that exploits radiating luminescence to excite nearby fluorophores by epifluorescence. We show that photons from bioluminescent bacteria radiate over mesoscopic distances and induce a red-shifted fluorescent emission from appropriate fluorophores in a manner distinct from bioluminescence resonance energy transfer. Our results characterizing fluorescence by unbound excitation from luminescence, both in vitro and in vivo, demonstrate how the resulting blue-to-red wavelength shift is both necessary and sufficient to yield contrast enhancement revealing mesoscopic proximity of luminescent and fluorescent probes in the context of living biological tissues. PMID:22615349

  6. Cerenkov Radiation as a New In Vivo Imaging Modality

    NASA Astrophysics Data System (ADS)

    Ackerman, Nicole; Ali, Rehan; Noll, J. Matt; Graves, Edward

    2011-04-01

    Čerenkov radiation has been used in particle detectors for years, but has recently been ``rediscovered'' by biologists working with radioactive isotopes. Čerenkov Light Imaging (CLI) can be done with CCD devices typically used for fluorescence or bioluminescence imaging. This provides the first opportunity for in vivo imaging of beta emitting isotopes, such as those used for radiation therapy. The GEANT4 simulation package has been used to simulate the properties and limitations of CLI. The simulation begins with the radioactive decay, generates the Čerenkov photons, propagates the optical light through tissue and other materials, and allows for different detection geometries. The simulation results are compared to in vivo and in vitro data taken in the Stanford Small Animal Imaging Core Facility.

  7. Nanoparticles that communicate in vivo to amplify tumour targeting

    NASA Astrophysics Data System (ADS)

    von Maltzahn, Geoffrey; Park, Ji-Ho; Lin, Kevin Y.; Singh, Neetu; Schwöppe, Christian; Mesters, Rolf; Berdel, Wolfgang E.; Ruoslahti, Erkki; Sailor, Michael J.; Bhatia, Sangeeta N.

    2011-07-01

    Nanomedicines have enormous potential to improve the precision of cancer therapy, yet our ability to efficiently home these materials to regions of disease in vivo remains very limited. Inspired by the ability of communication to improve targeting in biological systems, such as inflammatory-cell recruitment to sites of disease, we construct systems where synthetic biological and nanotechnological components communicate to amplify disease targeting in vivo. These systems are composed of ‘signalling’ modules (nanoparticles or engineered proteins) that target tumours and then locally activate the coagulation cascade to broadcast tumour location to clot-targeted ‘receiving’ nanoparticles in circulation that carry a diagnostic or therapeutic cargo, thereby amplifying their delivery. We show that communicating nanoparticle systems can be composed of multiple types of signalling and receiving modules, can transmit information through multiple molecular pathways in coagulation, can operate autonomously and can target over 40 times higher doses of chemotherapeutics to tumours than non-communicating controls.

  8. Assessing antibody pharmacokinetics in mice with in vivo imaging.

    PubMed

    Hoppin, Jack; Orcutt, Kelly Davis; Hesterman, Jacob Y; Silva, Matthew D; Cheng, Dengfeng; Lackas, Christian; Rusckowski, Mary

    2011-05-01

    Recent advances in small-animal molecular imaging instrumentation combined with well characterized antibody-labeling chemistry have enabled detailed in vivo measurements of antibody distribution in mouse models. This article reviews the strengths and limitations of in vivo antibody imaging methods with a focus on positron emission tomography and single-photon emission computed tomography and a brief discussion of the role of optical imaging in this application. A description of the basic principles behind the imaging techniques is provided along with a discussion of radiolabeling methods relevant to antibodies. Practical considerations of study design and execution are presented through a discussion of sensitivity and resolution tradeoffs for these techniques as defined by modality, signaling probe (isotope or fluorophore) selection, labeling method, and radiation dosimetry. Images and analysis results from a case study are presented with a discussion of output data content and relevant informatics gained with this approach to studying antibody pharmacokinetics. PMID:21317355

  9. Label-free oxygen-metabolic photoacoustic microscopy in vivo

    NASA Astrophysics Data System (ADS)

    Yao, Junjie; Maslov, Konstantin I.; Zhang, Yu; Xia, Younan; Wang, Lihong V.

    2011-07-01

    Almost all diseases, especially cancer and diabetes, manifest abnormal oxygen metabolism. Accurately measuring the metabolic rate of oxygen (MRO2) can be helpful for fundamental pathophysiological studies, and even early diagnosis and treatment of disease. Current techniques either lack high resolution or rely on exogenous contrast. Here, we propose label-free metabolic photoacoustic microscopy (mPAM) with small vessel resolution to noninvasively quantify MRO2 in vivo in absolute units. mPAM is the unique modality for simultaneously imaging all five anatomical, chemical, and fluid-dynamic parameters required for such quantification: tissue volume, vessel cross-section, concentration of hemoglobin, oxygen saturation of hemoglobin, and blood flow speed. Hyperthermia, cryotherapy, melanoma, and glioblastoma were longitudinally imaged in vivo. Counterintuitively, increased MRO2 does not necessarily cause hypoxia or increase oxygen extraction. In fact, early-stage cancer was found to be hyperoxic despite hypermetabolism.

  10. Applications of nuclear technologies for in-vivo elemental analysis

    SciTech Connect

    Cohn, S.H.; Ellis, K.J.; Vartsky, D.; Wielopolski, L.

    1982-01-01

    Measurement facilities developed, to date, include a unique whole-body-counter, (WBC); a total-body neutron-activation facility (TBNAA); and a partial-body activation facility (PBNAA). A variation of the prompt-gamma neutron-activation technique for measuring total-body nitrogen was developed to study body composition of cancer patients and the effect of nutritional regimens on the composition. These new techniques provide data in numerous clinical studies not previously amenable to investigation. The development and perfection of these techniques provide unique applications of radiation and radioisotopes to the early diagnosis of certain diseases and the evaluation of therapeutic programs. The PBNAA technique has been developed and calibrated for in-vivo measurement of metals. Development has gone forward on prompt-gamma neutron activation for the measurement of cadmium, x-ray fluorescence (XRF) for measurement of iron. Other techniques are being investigated for in-vivo measurement of metals such as silicon and beryllium.

  11. Nanoscale Imaging of Caveolin-1 Membrane Domains In Vivo

    PubMed Central

    Gabor, Kristin A.; Kim, Dahan; Kim, Carol H.; Hess, Samuel T.

    2015-01-01

    Light microscopy enables noninvasive imaging of fluorescent species in biological specimens, but resolution is generally limited by diffraction to ~200–250 nm. Many biological processes occur on smaller length scales, highlighting the importance of techniques that can image below the diffraction limit and provide valuable single-molecule information. In recent years, imaging techniques have been developed which can achieve resolution below the diffraction limit. Utilizing one such technique, fluorescence photoactivation localization microscopy (FPALM), we demonstrated its ability to construct super-resolution images from single molecules in a living zebrafish embryo, expanding the realm of previous super-resolution imaging to a living vertebrate organism. We imaged caveolin-1 in vivo, in living zebrafish embryos. Our results demonstrate the successful image acquisition of super-resolution images in a living vertebrate organism, opening several opportunities to answer more dynamic biological questions in vivo at the previously inaccessible nanoscale. PMID:25646724

  12. High-Resolution Multiphoton Imaging of Tumors In Vivo

    PubMed Central

    Wyckoff, Jeffrey; Gligorijevic, Bojana; Entenberg, David; Segall, Jeffrey; Condeelis, John

    2014-01-01

    Analysis of the individual steps in metastasis is crucial if insights at the molecular level are to be linked to the cell biology of cancer. A technical hurdle to achieving the analysis of the individual steps of metastasis is the fact that, at the gross level, tumors are heterogeneous in both animal models and patients. Human primary tumors show extensive variation in all properties ranging from growth and morphology of the tumor through tumor-cell density in the blood and formation and growth of metastases. Methods capable of the direct visualization and analysis of tumor-cell behavior at single-cell resolution in vivo have become crucial in advancing the understanding of mechanisms of metastasis, the definition of microenvironment, and the markers related to both. This article discusses the use of high-resolution multiphoton imaging of tumors (specifically breast tumors in mice) in vivo. PMID:21969629

  13. RNA nanotechnology for computer design and in vivo computation

    PubMed Central

    Qiu, Meikang; Khisamutdinov, Emil; Zhao, Zhengyi; Pan, Cheryl; Choi, Jeong-Woo; Leontis, Neocles B.; Guo, Peixuan

    2013-01-01

    Molecular-scale computing has been explored since 1989 owing to the foreseeable limitation of Moore's law for silicon-based computation devices. With the potential of massive parallelism, low energy consumption and capability of working in vivo, molecular-scale computing promises a new computational paradigm. Inspired by the concepts from the electronic computer, DNA computing has realized basic Boolean functions and has progressed into multi-layered circuits. Recently, RNA nanotechnology has emerged as an alternative approach. Owing to the newly discovered thermodynamic stability of a special RNA motif (Shu et al. 2011 Nat. Nanotechnol. 6, 658–667 (doi:10.1038/nnano.2011.105)), RNA nanoparticles are emerging as another promising medium for nanodevice and nanomedicine as well as molecular-scale computing. Like DNA, RNA sequences can be designed to form desired secondary structures in a straightforward manner, but RNA is structurally more versatile and more thermodynamically stable owing to its non-canonical base-pairing, tertiary interactions and base-stacking property. A 90-nucleotide RNA can exhibit 490 nanostructures, and its loops and tertiary architecture can serve as a mounting dovetail that eliminates the need for external linking dowels. Its enzymatic and fluorogenic activity creates diversity in computational design. Varieties of small RNA can work cooperatively, synergistically or antagonistically to carry out computational logic circuits. The riboswitch and enzymatic ribozyme activities and its special in vivo attributes offer a great potential for in vivo computation. Unique features in transcription, termination, self-assembly, self-processing and acid resistance enable in vivo production of RNA nanoparticles that harbour various regulators for intracellular manipulation. With all these advantages, RNA computation is promising, but it is still in its infancy. Many challenges still exist. Collaborations between RNA nanotechnologists and computer

  14. In vivo brain microdialysis: advances in neuropsychopharmacology and drug discovery

    PubMed Central

    Darvesh, Altaf S.; Carroll, Richard T.; Geldenhuys, Werner J.; Gudelsky, Gary A.; Klein, Jochen; Meshul, Charles K.; Van der Schyf, Cornelis J.

    2010-01-01

    Introduction Microdialysis is an important in vivo sampling technique, useful in the assay of extracellular tissue fluid. The technique has both pre-clinical and clinical applications but is most widely used in neuroscience. The in vivo microdialysis technique allows measurement of neurotransmitters such as acetycholine (ACh), the biogenic amines including dopamine (DA), norepinephrine (NE) and serotonin (5-HT), amino acids such as glutamate (Glu) and gamma aminobutyric acid (GABA), as well as the metabolites of the aforementioned neurotransmitters, and neuropeptides in neuronal extracellular fluid in discrete brain regions of laboratory animals such as rodents and non-human primates. Areas covered In this review we present a brief overview of the principles and procedures related to in vivo microdialysis and detail the use of this technique in the pre-clinical measurement of drugs designed to be used in the treatment of chemical addiction, neurodegenerative diseases such as Alzheimer’s disease (AD), Parkinson’s disease (PD) and as well as psychiatric disorders such as attention-deficit/hyperactivity disorder (ADHD) and schizophrenia. This review offers insight into the tremendous utility and versatility of this technique in pursuing neuropharmacological investigations as well its significant potential in rational drug discovery. Expert opinion In vivo microdialysis is an extremely versatile technique, routinely used in the neuropharmacological investigation of drugs used for the treatment of neurological disorders. This technique has been a boon in the elucidation of the neurochemical profile and mechanism of action of several classes of drugs especially their effects on neurotransmitter systems. The exploitation and development of this technique for drug discovery in the near future will enable investigational new drug candidates to be rapidly moved into the clinical trial stages and to market thus providing new successful therapies for neurological diseases

  15. In vivo method for the assessment of platelet accumulation

    SciTech Connect

    Smith, D.; Sanjar, S.; Herd, C.; Morley, J.

    1989-03-01

    Platelets labeled with 111-Indium have been injected intravenously into anesthetised guinea pigs, and intrathoracic content of 111-Indium-labeled platelets has been monitored continuously using a microcomputer-based system (AIMSplus). Dose-effect relationships have been described for ADP, collagen, and PAF, and effects of drugs upon selected responses illustrate the potential of this test system for routine in vivo screening of agents that may inhibit aggregation of platelets.

  16. Noninvasive in vivo determination of residual strains and stresses.

    PubMed

    Donmazov, Samir; Piskin, Senol; Pekkan, Kerem

    2015-06-01

    Vascular growth and remodeling during embryonic development are associated with blood flow and pressure induced stress distribution, in which residual strains and stresses play a central role. Residual strains are typically measured by performing in vitro tests on the excised vascular tissue. In this paper, we investigated the possibility of estimating residual strains and stresses using physiological pressure-radius data obtained through in vivo noninvasive measurement techniques, such as optical coherence tomography or ultrasound modalities. This analytical approach first tested with in vitro results using experimental data sets for three different arteries such as rabbit carotid artery, rabbit thoracic artery, and human carotid artery based on Fung's pseudostrain energy function and Delfino's exponential strain energy function (SEF). We also examined residual strains and stresses in the human swine iliac artery using the in vivo experimental ultrasound data sets corresponding to the systolic-to-diastolic region only. This allowed computation of the in vivo residual stress information for loading and unloading states separately. Residual strain parameters as well as the material parameters were successfully computed with high accuracy, where the relative errors are introduced in the range of 0-7.5%. Corresponding residual stress distributions demonstrated global errors all in acceptable ranges. A slight discrepancy was observed in the computed reduced axial force. Results of computations performed based on in vivo experimental data obtained from loading and unloading states of the artery exhibited alterations in material properties and residual strain parameters as well. Emerging noninvasive measurement techniques combined with the present analytical approach can be used to estimate residual strains and stresses in vascular tissues as a precursor for growth estimates. This approach is also validated with a finite element model of a general two-layered artery

  17. In vivo compartmental analysis of leukocytes in mouse lungs.

    PubMed

    Patel, Brijesh V; Tatham, Kate C; Wilson, Michael R; O'Dea, Kieran P; Takata, Masao

    2015-10-01

    The lung has a unique structure consisting of three functionally different compartments (alveolar, interstitial, and vascular) situated in an extreme proximity. Current methods to localize lung leukocytes using bronchoalveolar lavage and/or lung perfusion have significant limitations for determination of location and phenotype of leukocytes. Here we present a novel method using in vivo antibody labeling to enable accurate compartmental localization/quantification and phenotyping of mouse lung leukocytes. Anesthetized C57BL/6 mice received combined in vivo intravenous and intratracheal labeling with fluorophore-conjugated anti-CD45 antibodies, and lung single-cell suspensions were analyzed by flow cytometry. The combined in vivo intravenous and intratracheal CD45 labeling enabled robust separation of the alveolar, interstitial, and vascular compartments of the lung. In naive mice, the alveolar compartment consisted predominantly of resident alveolar macrophages. The interstitial compartment, gated by events negative for both intratracheal and intravenous CD45 staining, showed two conventional dendritic cell populations, as well as a Ly6C(lo) monocyte population. Expression levels of MHCII on these interstitial monocytes were much higher than on the vascular Ly6C(lo) monocyte populations. In mice exposed to acid aspiration-induced lung injury, this protocol also clearly distinguished the three lung compartments showing the dynamic trafficking of neutrophils and exudative monocytes across the lung compartments during inflammation and resolution. This simple in vivo dual-labeling technique substantially increases the accuracy and depth of lung flow cytometric analysis, facilitates a more comprehensive examination of lung leukocyte pools, and enables the investigation of previously poorly defined "interstitial" leukocyte populations during models of inflammatory lung diseases. PMID:26254421

  18. Intravital FRET: Probing Cellular and Tissue Function in Vivo.

    PubMed

    Radbruch, Helena; Bremer, Daniel; Mothes, Ronja; Günther, Robert; Rinnenthal, Jan Leo; Pohlan, Julian; Ulbricht, Carolin; Hauser, Anja E; Niesner, Raluca

    2015-01-01

    The development of intravital Förster Resonance Energy Transfer (FRET) is required to probe cellular and tissue function in the natural context: the living organism. Only in this way can biomedicine truly comprehend pathogenesis and develop effective therapeutic strategies. Here we demonstrate and discuss the advantages and pitfalls of two strategies to quantify FRET in vivo-ratiometrically and time-resolved by fluorescence lifetime imaging-and show their concrete application in the context of neuroinflammation in adult mice. PMID:26006244

  19. In vivo activity of aryl ozonides against Schistosoma species.

    PubMed

    Keiser, Jennifer; Ingram, Katrin; Vargas, Mireille; Chollet, Jacques; Wang, Xiaofang; Dong, Yuxiang; Vennerstrom, Jonathan L

    2012-02-01

    We evaluated the in vivo antischistosomal activities of 11 structurally diverse synthetic peroxides. Of all compounds tested, ozonide (1,2,4-trioxolane) OZ418 had the highest activity against adult Schistosoma mansoni, with total and female worm burden reductions of 80 and 90% (P < 0.05), respectively. Furthermore, treatment of S. haematobium-infected mice with OZ418 reduced the total worm burden by 86%. In conclusion, OZ418 is a promising antischistosomal lead compound. PMID:22106214

  20. IgA EGFR antibodies mediate tumour killing in vivo

    PubMed Central

    Boross, Peter; Lohse, Stefan; Nederend, Maaike; Jansen, Johannes Hendrik Marco; van Tetering, Geert; Dechant, Michael; Peipp, Matthias; Royle, Louise; Liew, Li Phing; Boon, Louis; van Rooijen, Nico; Bleeker, Wim K; Parren, Paul W H I; van de Winkel, Jan G J; Valerius, Thomas; Leusen, Jeanette H W

    2013-01-01

    Currently all approved anti-cancer therapeutic monoclonal antibodies (mAbs) are of the IgG isotype, which rely on Fcgamma receptors (FcγRs) to recruit cellular effector functions. In vitro studies showed that targeting of FcαRI (CD89) by bispecific antibodies (bsAbs) or recombinant IgA resulted in more effective elimination of tumour cells by myeloid effector cells than targeting of FcγR. Here we studied the in vivo anti-tumour activity of IgA EGFR antibodies generated using the variable sequences of the chimeric EGFR antibody cetuximab. Using FcαRI transgenic mice, we demonstrated significant in vivo anti-tumour activity of IgA2 EGFR against A431 cells in peritoneal and lung xenograft models, as well as against B16F10-EGFR cells in a lung metastasis model in immunocompetent mice. IgA2 EGFR was more effective than cetuximab in a short-term syngeneic peritoneal model using EGFR-transfected Ba/F3 target cells. The in vivo cytotoxic activity of IgA2 EGFR was mediated by macrophages and was significantly decreased in the absence of FcαRI. These results support the potential of targeting FcαRI for effective antibody therapy of cancer. The study reveals that IgA antibodies directed against EGFR and engaging Fcalpha receptor (FcαRI) on effector cells, have in vivo anti-cancer activity. These data support the development of novel immunotherapeutic strategies based on targeting FcαRI. PMID:23918228

  1. Advances in fiber optic sensors for in-vivo monitoring

    NASA Astrophysics Data System (ADS)

    Baldini, Francesco; Mignani, Anna G.

    1995-09-01

    Biomedical fiber-optic sensors are attractive for the measurement of both physical and chemical parameters as well as for spectral measurements directly performed on the patient. An overview of fiber-optic sensors for in vivo monitoring is given, with particular attention to the advantages that these sensors are able to offer in different fields of application such as cardiovascular and intensive care, angiology, gastroenterology, ophthalmology, oncology, neurology, dermatology, and dentistry.

  2. Targeted erythropoietin selectively stimulates red blood cell expansion in vivo.

    PubMed

    Burrill, Devin R; Vernet, Andyna; Collins, James J; Silver, Pamela A; Way, Jeffrey C

    2016-05-10

    The design of cell-targeted protein therapeutics can be informed by natural protein-protein interactions that use cooperative physical contacts to achieve cell type specificity. Here we applied this approach in vivo to the anemia drug erythropoietin (EPO), to direct its activity to EPO receptors (EPO-Rs) on red blood cell (RBC) precursors and prevent interaction with EPO-Rs on nonerythroid cells, such as platelets. Our engineered EPO molecule was mutated to weaken its affinity for EPO-R, but its avidity for RBC precursors was rescued via tethering to an antibody fragment that specifically binds the human RBC marker glycophorin A (huGYPA). We systematically tested the impact of these engineering steps on in vivo markers of efficacy, side effects, and pharmacokinetics. huGYPA transgenic mice dosed with targeted EPO exhibited elevated RBC levels, with only minimal platelet effects. This in vivo selectivity depended on the weakening EPO mutation, fusion to the RBC-specific antibody, and expression of huGYPA. The terminal plasma half-life of targeted EPO was ∼28.3 h in transgenic mice vs. ∼15.5 h in nontransgenic mice, indicating that huGYPA on mature RBCs acted as a significant drug sink but did not inhibit efficacy. In a therapeutic context, our targeting approach may allow higher restorative doses of EPO without platelet-mediated side effects, and also may improve drug pharmacokinetics. These results demonstrate how rational drug design can improve in vivo specificity, with potential application to diverse protein therapeutics. PMID:27114509

  3. RNA nanotechnology for computer design and in vivo computation.

    PubMed

    Qiu, Meikang; Khisamutdinov, Emil; Zhao, Zhengyi; Pan, Cheryl; Choi, Jeong-Woo; Leontis, Neocles B; Guo, Peixuan

    2013-10-13

    Molecular-scale computing has been explored since 1989 owing to the foreseeable limitation of Moore's law for silicon-based computation devices. With the potential of massive parallelism, low energy consumption and capability of working in vivo, molecular-scale computing promises a new computational paradigm. Inspired by the concepts from the electronic computer, DNA computing has realized basic Boolean functions and has progressed into multi-layered circuits. Recently, RNA nanotechnology has emerged as an alternative approach. Owing to the newly discovered thermodynamic stability of a special RNA motif (Shu et al. 2011 Nat. Nanotechnol. 6, 658-667 (doi:10.1038/nnano.2011.105)), RNA nanoparticles are emerging as another promising medium for nanodevice and nanomedicine as well as molecular-scale computing. Like DNA, RNA sequences can be designed to form desired secondary structures in a straightforward manner, but RNA is structurally more versatile and more thermodynamically stable owing to its non-canonical base-pairing, tertiary interactions and base-stacking property. A 90-nucleotide RNA can exhibit 4⁹⁰ nanostructures, and its loops and tertiary architecture can serve as a mounting dovetail that eliminates the need for external linking dowels. Its enzymatic and fluorogenic activity creates diversity in computational design. Varieties of small RNA can work cooperatively, synergistically or antagonistically to carry out computational logic circuits. The riboswitch and enzymatic ribozyme activities and its special in vivo attributes offer a great potential for in vivo computation. Unique features in transcription, termination, self-assembly, self-processing and acid resistance enable in vivo production of RNA nanoparticles that harbour various regulators for intracellular manipulation. With all these advantages, RNA computation is promising, but it is still in its infancy. Many challenges still exist. Collaborations between RNA nanotechnologists and computer

  4. Photoacoustic monitoring and imaging of blood vessel size in vivo

    NASA Astrophysics Data System (ADS)

    Kolkman, Roy G. M.; Klaessens, John H. G. M.; Hondebrink, Erwin; Hopman, Jeroen C. W.; Steenbergen, Wiendelt; van Leeuwen, Ton G.; Thijssen, Johan M.; de Mul, Frits F. M.

    2003-06-01

    A double-ring photoacoustic sensor to image and monitor blood content in tissue has been developed. This sensor has a very small opening angle. Using this sensor we are able to image artifical blood vessels, as well as vessels in a rabbit ear. Furthermore, the feasibility of in vivo imaging is demonstrated with a photoacoustic reconstruction of the joining of two palmar veins a few centimeter proximal to the wrist in a human arm.

  5. Grueneisen relaxation photoacoustic microscopy in vivo (Conference Presentation)

    NASA Astrophysics Data System (ADS)

    Ma, Jun; Shi, Junhui; Hai, Pengfei; Zhou, Yong; Wang, Lihong V.

    2016-03-01

    Optical-resolution photoacoustic microscopy (OR-PAM) can achieve submicron lateral resolution by tightly focusing the excitation light, while the axial resolution is still limited by the frequency bandwidth of the ultrasonic transducer. The Grueneisen relaxation effect, in which the Grueneisen parameter changes within the thermal relaxation time following a laser impulse heating, can provide excellent axial resolution due to its optical sectioning property. Based on this effect, Grueneisen relaxation photoacoustic microscopy (GR-PAM) was developed and demonstrated ex vivo. Here, we present for the first time in vivo imaging of mouse brains with improved axial resolution based on GR-PAM. An intensity-modulated continuous-wave (CW) 532 nm laser thermally heated the in-focus absorber. Another 532 nm pulsed laser, which is aligned confocally with the CW laser, generated the photoacoustic (PA) signal from the absorber. The difference between the amplitudes of the photoacoustic signals with and without heating was used for image reconstruction. The achieved axial resolution is ~12.5 µm, which is fivefold better than the acoustically determined value for a 20 MHz-bandwidth ultrasound transducer. The system was demonstrated by imaging a blood-filled tube ex vivo and blood vessels of mouse brains in vivo. The blood-filled tube diameter obtained from the PA image by GR-PAM is 105 µm, which is much closer to its actual diameter (100 µm) than the value from conventional OR-PAM (160 µm). This axial resolution improvement was further validated in imaging mouse brains in vivo, and yielded significantly narrower axial profiles of the vessels. This in vivo demonstration of imaging by GR-PAM might inspire more applications in PA biomedical imaging and sensing.

  6. In vivo real-time volumetric synthetic aperture ultrasound imaging

    NASA Astrophysics Data System (ADS)

    Bouzari, Hamed; Rasmussen, Morten F.; Brandt, Andreas H.; Stuart, Matthias B.; Nikolov, Svetoslav; Jensen, Jørgen A.

    2015-03-01

    Synthetic aperture (SA) imaging can be used to achieve real-time volumetric ultrasound imaging using 2-D array transducers. The sensitivity of SA imaging is improved by maximizing the acoustic output, but one must consider the limitations of an ultrasound system, both technical and biological. This paper investigates the in vivo applicability and sensitivity of volumetric SA imaging. Utilizing the transmit events to generate a set of virtual point sources, a frame rate of 25 Hz for a 90° × 90° field-of-view was achieved. data were obtained using a 3.5 MHz 32 × 32 elements 2-D phased array transducer connected to the experimental scanner (SARUS). Proper scaling is applied to the excitation signal such that intensity levels are in compliance with the U.S. Food and Drug Administration regulations for in vivo ultrasound imaging. The measured Mechanical Index and spatial-peak-temporal-average intensity for parallel beam-forming (PB) are 0.83 and 377.5mW/cm2, and for SA are 0.48 and 329.5mW/cm2. A human kidney was volumetrically imaged with SA and PB techniques simultaneously. Two radiologists for evaluation of the volumetric SA were consulted by means of a questionnaire on the level of details perceivable in the beam-formed images. The comparison was against PB based on the in vivo data. The feedback from the domain experts indicates that volumetric SA images internal body structures with a better contrast resolution compared to PB at all positions in the entire imaged volume. Furthermore, the autocovariance of a homogeneous area in the in vivo SA data, had 23.5% smaller width at the half of its maximum value compared to PB.

  7. Toxicity of methyldopa (Aldomet) to mouse neuroblastoma cells in vivo.

    PubMed

    Chelmicka-Schorr, E; Sportiello, M G; Otten, G R; Arnason, B G

    1983-08-01

    The adrenergic blocking agent methyldopa (Aldomet) is toxic to C-1300 neuroblastoma cells in vivo. Four injections of Aldomet at a dose of 7.5 mg/injection were given over a period of 24 hr to C-1300 neuroblastoma-bearing mice. This treatment killed a significant proportion of the C-1300 neuroblastoma cells. Flow cytometric data suggest that sensitivity of tumor cells to Aldomet is not related to the cell cycle. PMID:6861122

  8. Measuring In Vivo Free Radical Production by the Outer Retina

    PubMed Central

    Berkowitz, Bruce A.; Bredell, Bryce X.; Davis, Christopher; Samardzija, Marijana; Grimm, Christian; Roberts, Robin

    2015-01-01

    Purpose Excessive and continuously produced free radicals in the outer retina are implicated in retinal aging and the pathogenesis of sight-threatening retinopathies, yet measuring outer retinal oxidative stress in vivo remains a challenge. Here, we test the hypothesis that continuously produced paramagnetic free radicals from the outer retina can be measured in vivo using high-resolution (22-μm axial resolution) 1/T1magnetic resonance imaging (MRI) without and with a confirmatory quench (quench-assisted MRI). Methods Low-dose sodium iodate–treated and diabetic C57Bl6/J mice (and their controls), and rod-dominated (129S6) or cone-only R91W;Nrl−/− mice were studied. In dark-adapted groups, 1/T1 was mapped transretinally in vivo without or with (1) the antioxidant combination of methylene blue (MB) and α-lipoic acid (LPA), or (2) light exposure; in subgroups, retinal superoxide production was measured ex vivo (lucigenin). Results In the sodium iodate model, retinal superoxide production and outer retina-specific 1/T1 values were both significantly greater than normal and corrected to baseline with MB+LPA therapy. Nondiabetic mice at two ages and 1.2-month diabetic mice (before the appearance of oxidative stress) had similar transretinal 1/T1 profiles. By 2.3 months of diabetes, only outer retinal 1/T1 values were significantly greater than normal and were corrected to baseline with MB+LPA therapy. In mice with healthy photoreceptors, a light quench caused 1/T1 of rods, but not cones, to significantly decrease from their values in the dark. Conclusions Quench-assisted MRI is a feasible method for noninvasively measuring normal and pathologic production of free radicals in photoreceptors/RPE in vivo. PMID:26670830

  9. Multiplexed aberration measurement for deep tissue imaging in vivo

    PubMed Central

    Wang, Chen; Liu, Rui; Milkie, Daniel E.; Sun, Wenzhi; Tan, Zhongchao; Kerlin, Aaron; Chen, Tsai-Wen; Kim, Douglas S.; Ji, Na

    2014-01-01

    We describe a multiplexed aberration measurement method that modulates the intensity or phase of light rays at multiple pupil segments in parallel to determine their phase gradients. Applicable to fluorescent-protein-labeled structures of arbitrary complexity, it allows us to obtain diffraction-limited resolution in various samples in vivo. For the strongly scattering mouse brain, a single aberration correction improves structural and functional imaging of fine neuronal processes over a large imaging volume. PMID:25128976

  10. Capacitive Extensometer Particularly Suited for Measuring in Vivo Bone Strain

    NASA Technical Reports Server (NTRS)

    Perusek, Gail P. (Inventor)

    2000-01-01

    The present invention provides for in vivo measurements of the principal strain magnitudes and directions, and maximum shear strain that occurs in a material, such as human bone, when it is loaded (or subjected to a load). In one embodiment the invention includes a capacitive delta extensometer arranged with six sensors in a three piece configuration, with each sensor of each pair spaced apart from each other by 120 degrees.

  11. In vivo EMG biofeedback in violin and viola pedagogy.

    PubMed

    LeVine, W R; Irvine, J K

    1984-06-01

    In vivo EMG biofeedback was found to be an effective pedagogical tool for removing unwanted left-hand tension in nine violin and viola players. Improvement occurred rapidly and persisted throughout a 5-month follow-up period. Further studies will be necessary to assess the effect of biofeedback independent of placebo effects. The brevity of the method and the magnitude of improvement warrant further investigation. PMID:6509108

  12. Interferon induces natural killer cell blastogenesis in vivo

    NASA Technical Reports Server (NTRS)

    Biron, C. A.; Sonnenfeld, G.; Welsh, R. M.

    1984-01-01

    Interferon (IFN), types beta and gamma, and IFN inducers polyinosinic-polycytidylic acid and lymphocytic choriomeningitis virus, all stimulated the generation of blast-natural killer (NK) cells in mouse spleens, Blast-NK cells were characterized on the basis of size, 3H-thymidine uptake, and NK cell markers These data indicate that in addition to augmenting NK cell-mediated lysis, IFN may regulate NK cell proliferation in vivo.

  13. Studying tumor metastasis by in vivo imaging and flow cytometer

    NASA Astrophysics Data System (ADS)

    Wei, Xunbin; Guo, Jin; Liu, Guangda; Li, Yan; Chen, Yun; Zhang, Li; Tan, Yuan; Chen, Tong; Gu, Zhenqin; Wang, Chen

    2009-02-01

    Liver cancer is one of the most common malignancies in the world, with approximately 1,000,000 cases reported every year. This ranges from 15,000 cases in the United States to more than a 250,000 in China. About 80% of people with primary liver cancer are male. Although two-thirds of people have advanced liver disease when they seek medical help, one third of the patients have cancer that has not progressed beyond the liver. Primary liver cancer (hepatocellular carcinoma, or HCC) is associated with liver cirrhosis 60-80% of the time. HCC may metastasize to the lung, bones, kidney, and many other organs. Surgical resection, liver transplantation, chemotherapy and radiation therapy are the foundation of current HCC therapies. However the outcomes are poor-the survival rate is almost zero for metastatic HCC patients. Molecular mechanisms of HCC metastasis need to be understood better and new therapies must be developed to selectively target to unique characteristics of HCC cell growth and metastasis. We have developed the "in vivo microscopy" to study the mechanisms that govern liver tumor cell spread through the microenvironment in vivo in real-time confocal near-infrared fluorescence imaging. A recently developed "in vivo flow cytometer" and optical imaging are used to assess liver tumor cell spreading and the circulation kinetics of liver tumor cells. A real-time quantitative monitoring of circulating liver tumor cells by the in vivo flow cytometer will be useful to assess the effectiveness of the potential therapeutic interventions.

  14. Exploring the case for assisted dying in the UK.

    PubMed

    Haigh, Carol

    This article discusses the concepts of euthanasia, assisted suicide and physician-assisted suicide, under the umbrella term of assisted dying, from a pro-assisted dying perspective. It outlines the key principles underpinning the debate around assisted dying and refutes the main arguments put forward by those opposing legalisation of assisted dying in the UK. PMID:22272538

  15. Virulence of an emerging respiratory pathogen, genus Pandoraea, in vivo and its interactions with lung epithelial cells.

    PubMed

    Costello, Anne; Herbert, Gillian; Fabunmi, Lydia; Schaffer, Kirsten; Kavanagh, Kevin A; Caraher, Emma M; Callaghan, Máire; McClean, Siobhán

    2011-03-01

    Pandoraea species have emerged as opportunistic pathogens among cystic fibrosis (CF) and non-CF patients. Pandoraea pulmonicola is the predominant Pandoraea species among Irish CF patients. The objective of this study was to investigate the pathogenicity and potential mechanisms of virulence of Irish P. pulmonicola isolates and strains from other Pandoraea species. Three patients from whom the P. pulmonicola isolates were isolated have since died. The in vivo virulence of these and other Pandoraea strains was examined by determining the ability to kill Galleria mellonella larvae. The P. pulmonicola strains generally were the most virulent of the species tested, with three showing a comparable or greater level of virulence in vivo relative to another CF pathogen, Burkholderia cenocepacia, whilst strains from two other species, Pandoraea apista and Pandoraea pnomenusa, were considerably less virulent. For all Pandoraea species, whole cells were required for larval killing, as cell-free supernatants had little effect on larval survival. Overall, invasive Pandoraea strains showed comparable invasion of two independent lung epithelial cell lines, irrespective of whether they had a CF phenotype. Pandoraea strains were also capable of translocation across polarized lung epithelial cell monolayers. Although protease secretion was a common characteristic across the genus, it is unlikely to be involved in pathogenesis. In conclusion, whilst multiple mechanisms of pathogenicity may exist across the genus Pandoraea, it appears that lung cell invasion and translocation contribute to the virulence of P. pulmonicola strains. PMID:21127160

  16. The slow Wallerian degeneration gene in vivo protects motor axons but not their cell bodies after avulsion and neonatal axotomy.

    PubMed

    Adalbert, Robert; Nógrádi, Antal; Szabó, András; Coleman, Michael P

    2006-10-01

    The slow Wallerian degeneration gene (Wld(S)) delays Wallerian degeneration and axon pathology for several weeks in mice and rats. Interestingly, neuronal cell death is also delayed in some in vivo models, most strikingly in the progressive motoneuronopathy mouse. Here, we tested the hypothesis that Wld(S) has a direct protective effect on motoneurone cell bodies in vivo. Cell death was induced in rat L4 motoneurones by intravertebral avulsion of the corresponding ventral roots. This simultaneously removed most of the motor axon, minimizing the possibility that the protective effect toward axons could rescue cell bodies secondarily. There was no significant difference between the survival of motoneurones in control and Wld(S) rats, suggesting that the Wld(S) gene has no direct protective effect on cell bodies. We also tested for any delay in apoptotic motoneurone death following neonatal nerve injury in Wld(S) rats and found that, unlike Wld(S) mice, Wld(S) rats show no delay in cell death. However, the corresponding distal axons were preserved, confirming that motoneurone cell bodies and motor axons die by different mechanisms. Thus, Wld(S) does not directly prevent death of motoneurone cell bodies. It follows that the protection of neuronal cell bodies observed in several disease and injury models where axons or significant axonal stumps remain is most probably secondary to axonal protection. PMID:17074042

  17. Toxoplasma gondii: in vivo and in vitro studies of a mutant resistant to arprinocid-N-oxide.

    PubMed

    Pfefferkorn, E R; Eckel, M E; McAdams, E

    1988-04-01

    The anticoccidial drug arprinocid and arprinocid-N-oxide, a metabolite produced in vivo, blocked the growth of Toxoplasma gondii in human fibroblasts. The more potent arprinocid-N-oxide inhibited growth by 50% at 20 ng/ml while arprinocid inhibited at 2 micrograms/ml. For both drugs, the host cell was less sensitive than was the parasite. Hypoxanthine did not reverse the antitoxoplasma activity of either drug. We isolated a parasite mutant, R-AnoR-1 that was 16- to 20-fold more resistant to arprinocid-N-oxide than was the wild type RH T. gondii. This mutant was not resistant to arprinocid in vitro. Arprinocid in a daily oral dose of 136 micrograms regularly protected mice against an otherwise fatal infection with RH T. gondii and 55 micrograms had some protective effect. However, all mice infected with R-AnoR-1 and treated with 360 micrograms arprinocid per day died. Since this mutant is fully sensitive to arprinocid, the form of the drug that is therapeutically active in vivo cannot be arprinocid and is likely to be arprinocid-N-oxide. PMID:3350108

  18. Diploic venous anatomy studied in-vivo by MRI.

    PubMed

    Jivraj, Khalil; Bhargava, Ravi; Aronyk, Keith; Quateen, Ahmed; Walji, Anil

    2009-04-01

    Calvarial diploic venous anatomy has been studied post-mortem, but few studies have addressed these venous structures in-vivo. Previous work in our laboratory has shown that intraosseous infusion through the skull diploic space near the diploic veins in animals and humans does access the superior sagittal sinus and the systemic venous system. We developed a volumetric method of imaging the diploic veins in-vivo using MRI, intravenous gadolinium, and digital subtraction to provide for three-dimensional depiction and exact localization of these veins. We hypothesized that this technique would allow for an assessment of the probability of existence, distribution, and concentration of diploic veins in the skull. We scanned 31 neurosurgical patients, and were able to create 3D diploic venous maps in 74% of them. These maps were processed using Adobe Photoshop CS2. Mathworks MatLab 6.5, once customized, counted the number of pixels occupied by the diploic veins in the processed image. The probability of veins was highest in the occipital regions (100%). The inferior occipital (4.1%) and posterior parietal (4.1%) regions had the highest concentrations of diploic veins. Digital subtraction venography using a volumetric MRI sequence can demonstrate the diploic veins in-vivo. The inferior occipital region may be the best area for an intraosseous infusion device because it has the greatest likelihood of containing a vein and also has the highest concentration of veins. PMID:19173254

  19. HLA-B27/microbial mimicry: an in vivo analysis.

    PubMed Central

    Kapasi, K; Chui, B; Inman, R D

    1992-01-01

    The association between three major spondyloarthritic diseases, ankylosing spondylitis, Reiter's syndrome, and reactive arthritis, and the major histocompatibility complex (MHC) class 1 antigen HLA-B27 is well documented. The hypothesis of cross-reactivity between HLA-B27 and the antecedent infection-causing Gram-negative pathogens such as Salmonella, Shigella and Yersinia has been suggested by in vitro studies employing monoclonal antibodies. We have examined the possibility of such cross-reactivity in vivo using various rabbit immune sera and patient sera as the source of cross-reacting antibody. Mouse L cells were transfected with HLA-A3 or HLA-B27 and used as a source of antigen. Western blot analysis employing denatured antigen, FACS analysis employing native antigen and immunoprecipitation studies were undertaken to detect cross-reacting antibodies generated in vivo to HLA-B27 antigen. Antibodies generated in vivo by infection in patients or immunization in animals against arthritogenic bacteria did not demonstrate any cross-reactivity with HLA-B27 by any of the methods used. As defined by the humoral immune response, molecular mimicry appears unlikely to explain the role of B27 in the pathogenesis of reactive arthritis. Images Figure 2 Figure 3 Figure 6 PMID:1478690

  20. Infrared neural stimulation of human spinal nerve roots in vivo

    PubMed Central

    Cayce, Jonathan M.; Wells, Jonathon D.; Malphrus, Jonathan D.; Kao, Chris; Thomsen, Sharon; Tulipan, Noel B.; Konrad, Peter E.; Jansen, E. Duco; Mahadevan-Jansen, Anita

    2015-01-01

    Abstract. Infrared neural stimulation (INS) is a neurostimulation modality that uses pulsed infrared light to evoke artifact-free, spatially precise neural activity with a noncontact interface; however, the technique has not been demonstrated in humans. The objective of this study is to demonstrate the safety and efficacy of INS in humans in vivo. The feasibility of INS in humans was assessed in patients (n=7) undergoing selective dorsal root rhizotomy, where hyperactive dorsal roots, identified for transection, were stimulated in vivo with INS on two to three sites per nerve with electromyogram recordings acquired throughout the stimulation. The stimulated dorsal root was removed and histology was performed to determine thermal damage thresholds of INS. Threshold activation of human dorsal rootlets occurred in 63% of nerves for radiant exposures between 0.53 and 1.23  J/cm2. In all cases, only one or two monitored muscle groups were activated from INS stimulation of a hyperactive spinal root identified by electrical stimulation. Thermal damage was first noted at 1.09  J/cm2 and a 2∶1 safety ratio was identified. These findings demonstrate the success of INS as a fresh approach for activating human nerves in vivo and providing the necessary safety data needed to pursue clinically driven therapeutic and diagnostic applications of INS in humans. PMID:26157986

  1. In vivo degeneration and the fate of inorganic nanoparticles.

    PubMed

    Feliu, Neus; Docter, Dominic; Heine, Markus; Del Pino, Pablo; Ashraf, Sumaira; Kolosnjaj-Tabi, Jelena; Macchiarini, Paolo; Nielsen, Peter; Alloyeau, Damien; Gazeau, Florence; Stauber, Roland H; Parak, Wolfgang J

    2016-05-01

    What happens to inorganic nanoparticles (NPs), such as plasmonic gold or silver, superparamagnetic iron oxide, or fluorescent quantum dot NPs after they have been administrated to a living being? This review discusses the integrity, biodistribution, and fate of NPs after in vivo administration. The hybrid nature of the NPs is described, conceptually divided into the inorganic core, the engineered surface coating comprising of the ligand shell and optionally also bio-conjugates, and the corona of adsorbed biological molecules. Empirical evidence shows that all of these three compounds may degrade individually in vivo and can drastically modify the life cycle and biodistribution of the whole heterostructure. Thus, the NPs may be decomposed into different parts, whose biodistribution and fate would need to be analyzed individually. Multiple labeling and quantification strategies for such a purpose will be discussed. All reviewed data indicate that NPs in vivo should no longer be considered as homogeneous entities, but should be seen as inorganic/organic/biological nano-hybrids with complex and intricately linked distribution and degradation pathways. PMID:26862602

  2. Imaging of gene expression in vivo with photoacoustic tomography

    NASA Astrophysics Data System (ADS)

    Li, Li; Zemp, Roger J.; Lungu, Gina; Stoica, George; Wang, Lihong V.

    2006-02-01

    In the post-genomic era, there is an increasing interest in visualizing the expression of functional genes in vivo. With the assistance of the reporter gene technique, various imaging modalities have been adopted for this purpose. In vivo gene expression imaging promises to provide biologists with a powerful tool for deepening our understanding of developmental biology, expanding our knowledge of the genetic basis of disease, and advancing the development of medicine. In this paper, we demonstrate the feasibility of imaging gene expression with photoacoustic imaging, which offers unique absorption contrast with ultrasonic resolution in vivo. We mark tumors in rats with the lacZ reporter gene. The lacZ gene encodes an enzyme β-galactosidase, which yields a dark blue product when acting on a colorimetric assay called X-gal. Photoacoustic tomography at 650nm clearly visualizes the presence of this blue product. The spectroscopic method can also potentially improve specificity. Considering how many staining methods are used in traditional biology, we believe that photoacoustic techniques will revolutionize the field of molecular imaging. The further development of reporter gene systems with high absorbing products in the NIR region is needed.

  3. Structural imprints in vivo decode RNA regulatory mechanisms

    PubMed Central

    Spitale, Robert C.; Flynn, Ryan A.; Zhang, Qiangfeng Cliff; Crisalli, Pete; Lee, Byron; Jung, Jong-Wha; Kuchelmeister, Hannes Y.; Batista, Pedro J.; Torre, Eduardo A.; Kool, Eric T.; Chang, Howard Y.

    2015-01-01

    Visualizing the physical basis for molecular behavior inside living cells is a grand challenge in biology. RNAs are central to biological regulation, and RNA’s ability to adopt specific structures intimately controls every step of the gene expression program1. However, our understanding of physiological RNA structures is limited; current in vivo RNA structure profiles view only two of four nucleotides that make up RNA2,3. Here we present a novel biochemical approach, In Vivo Click SHAPE (icSHAPE), that enables the first global view of RNA secondary structures of all four bases in living cells. icSHAPE of mouse embryonic stem cell transcriptome versus purified RNA folded in vitro shows that the structural dynamics of RNA in the cellular environment distinguishes different classes of RNAs and regulatory elements. Structural signatures at translational start sites and ribosome pause sites are conserved from in vitro, suggesting that these RNA elements are programmed by sequence. In contrast, focal structural rearrangements in vivo reveal precise interfaces of RNA with RNA binding proteins or RNA modification sites that are consistent with atomic-resolution structural data. Such dynamic structural footprints enable accurate prediction of RNA-protein interactions and N6-methyladenosine (m6A) modification genome-wide. These results open the door for structural genomics of RNA in living cells and reveal key physiological structures controlling gene expression. PMID:25799993

  4. Nanosensor aided photoacoustic measurement of pH in vivo

    NASA Astrophysics Data System (ADS)

    Ray, Aniruddha; Yoon, Hyung Ki; Kopelman, Raoul; Wang, Xueding

    2013-03-01

    pH plays a critical role in many aspects of cell and tissues physiology. Lower pH is also a typical characteristic of arthritic joints and tumor tissues. These pH anomalies are also exploited in different drug delivery mechanisms. Here we present, a new method of pH sensing in vivo using spectroscopic photoacoustic measurements facilitated by pH sensitive nanosensors. The nanosensors consist of Seminaphtharhodafluor (SNARF), a pH sensitive dye, encapsulated in a specially designed polyacrylamide hydrogel matrix with a hydrophobic core. The photoacoustic intensity ratio between the excitation wavelengths of 585nm and 565nm increases in the pH range from 6.0 to 8.0 and is used to determine the pH of the local environment. These nanosensors are biodegradable, biocompatible, have a long plasma lifetime and can be targeted to any type of cells or tissues by surface modification using proper targeting moieties. The encapsulation of the dye prevents the interaction of the dye with proteins in plasma and also reduces the dye degradation. The SNARF dye in its free form loses 90% of its absorbance in presence of albumin, a protein found in abundance in plasma, and this has severely limited its adaptation to in vivo environments. In comparison, the SNARF nanosensors lose only 16% of their absorbance in the same environment. We employ these nanosensors to demonstrate the feasibility of pH sensing in vivo through photoacoustic measurements on a rat joint model.

  5. Organelle acidification negatively regulates vacuole membrane fusion in vivo

    PubMed Central

    Desfougères, Yann; Vavassori, Stefano; Rompf, Maria; Gerasimaite, Ruta; Mayer, Andreas

    2016-01-01

    The V-ATPase is a proton pump consisting of a membrane-integral V0 sector and a peripheral V1 sector, which carries the ATPase activity. In vitro studies of yeast vacuole fusion and evidence from worms, flies, zebrafish and mice suggested that V0 interacts with the SNARE machinery for membrane fusion, that it promotes the induction of hemifusion and that this activity requires physical presence of V0 rather than its proton pump activity. A recent in vivo study in yeast has challenged these interpretations, concluding that fusion required solely lumenal acidification but not the V0 sector itself. Here, we identify the reasons for this discrepancy and reconcile it. We find that acute pharmacological or physiological inhibition of V-ATPase pump activity de-acidifies the vacuole lumen in living yeast cells within minutes. Time-lapse microscopy revealed that de-acidification induces vacuole fusion rather than inhibiting it. Cells expressing mutated V0 subunits that maintain vacuolar acidity were blocked in this fusion. Thus, proton pump activity of the V-ATPase negatively regulates vacuole fusion in vivo. Vacuole fusion in vivo does, however, require physical presence of a fusion-competent V0 sector. PMID:27363625

  6. A crystal-clear zebrafish for in vivo imaging

    PubMed Central

    Antinucci, Paride; Hindges, Robert

    2016-01-01

    The larval zebrafish (Danio rerio) is an excellent vertebrate model for in vivo imaging of biological phenomena at subcellular, cellular and systems levels. However, the optical accessibility of highly pigmented tissues, like the eyes, is limited even in this animal model. Typical strategies to improve the transparency of zebrafish larvae require the use of either highly toxic chemical compounds (e.g. 1-phenyl-2-thiourea, PTU) or pigmentation mutant strains (e.g. casper mutant). To date none of these strategies produce normally behaving larvae that are transparent in both the body and the eyes. Here we present crystal, an optically clear zebrafish mutant obtained by combining different viable mutations affecting skin pigmentation. Compared to the previously described combinatorial mutant casper, the crystal mutant lacks pigmentation also in the retinal pigment epithelium, therefore enabling optical access to the eyes. Unlike PTU-treated animals, crystal larvae are able to perform visually guided behaviours, such as the optomotor response, as efficiently as wild type larvae. To validate the in vivo application of crystal larvae, we performed whole-brain light-sheet imaging and two-photon calcium imaging of neural activity in the retina. In conclusion, this novel combinatorial pigmentation mutant represents an ideal vertebrate tool for completely unobstructed structural and functional in vivo investigations of biological processes, particularly when imaging tissues inside or between the eyes. PMID:27381182

  7. A Statistical Model for In Vivo Neuronal Dynamics

    PubMed Central

    Surace, Simone Carlo; Pfister, Jean-Pascal

    2015-01-01

    Single neuron models have a long tradition in computational neuroscience. Detailed biophysical models such as the Hodgkin-Huxley model as well as simplified neuron models such as the class of integrate-and-fire models relate the input current to the membrane potential of the neuron. Those types of models have been extensively fitted to in vitro data where the input current is controlled. Those models are however of little use when it comes to characterize intracellular in vivo recordings since the input to the neuron is not known. Here we propose a novel single neuron model that characterizes the statistical properties of in vivo recordings. More specifically, we propose a stochastic process where the subthreshold membrane potential follows a Gaussian process and the spike emission intensity depends nonlinearly on the membrane potential as well as the spiking history. We first show that the model has a rich dynamical repertoire since it can capture arbitrary subthreshold autocovariance functions, firing-rate adaptations as well as arbitrary shapes of the action potential. We then show that this model can be efficiently fitted to data without overfitting. We finally show that this model can be used to characterize and therefore precisely compare various intracellular in vivo recordings from different animals and experimental conditions. PMID:26571371

  8. A crystal-clear zebrafish for in vivo imaging.

    PubMed

    Antinucci, Paride; Hindges, Robert

    2016-01-01

    The larval zebrafish (Danio rerio) is an excellent vertebrate model for in vivo imaging of biological phenomena at subcellular, cellular and systems levels. However, the optical accessibility of highly pigmented tissues, like the eyes, is limited even in this animal model. Typical strategies to improve the transparency of zebrafish larvae require the use of either highly toxic chemical compounds (e.g. 1-phenyl-2-thiourea, PTU) or pigmentation mutant strains (e.g. casper mutant). To date none of these strategies produce normally behaving larvae that are transparent in both the body and the eyes. Here we present crystal, an optically clear zebrafish mutant obtained by combining different viable mutations affecting skin pigmentation. Compared to the previously described combinatorial mutant casper, the crystal mutant lacks pigmentation also in the retinal pigment epithelium, therefore enabling optical access to the eyes. Unlike PTU-treated animals, crystal larvae are able to perform visually guided behaviours, such as the optomotor response, as efficiently as wild type larvae. To validate the in vivo application of crystal larvae, we performed whole-brain light-sheet imaging and two-photon calcium imaging of neural activity in the retina. In conclusion, this novel combinatorial pigmentation mutant represents an ideal vertebrate tool for completely unobstructed structural and functional in vivo investigations of biological processes, particularly when imaging tissues inside or between the eyes. PMID:27381182

  9. Simultaneous in vivo positron emission tomography and magnetic resonance imaging.

    PubMed

    Catana, Ciprian; Procissi, Daniel; Wu, Yibao; Judenhofer, Martin S; Qi, Jinyi; Pichler, Bernd J; Jacobs, Russell E; Cherry, Simon R

    2008-03-11

    Positron emission tomography (PET) and magnetic resonance imaging (MRI) are widely used in vivo imaging technologies with both clinical and biomedical research applications. The strengths of MRI include high-resolution, high-contrast morphologic imaging of soft tissues; the ability to image physiologic parameters such as diffusion and changes in oxygenation level resulting from neuronal stimulation; and the measurement of metabolites using chemical shift imaging. PET images the distribution of biologically targeted radiotracers with high sensitivity, but images generally lack anatomic context and are of lower spatial resolution. Integration of these technologies permits the acquisition of temporally correlated data showing the distribution of PET radiotracers and MRI contrast agents or MR-detectable metabolites, with registration to the underlying anatomy. An MRI-compatible PET scanner has been built for biomedical research applications that allows data from both modalities to be acquired simultaneously. Experiments demonstrate no effect of the MRI system on the spatial resolution of the PET system and <10% reduction in the fraction of radioactive decay events detected by the PET scanner inside the MRI. The signal-to-noise ratio and uniformity of the MR images, with the exception of one particular pulse sequence, were little affected by the presence of the PET scanner. In vivo simultaneous PET and MRI studies were performed in mice. Proof-of-principle in vivo MR spectroscopy and functional MRI experiments were also demonstrated with the combined scanner. PMID:18319342

  10. High altitude impairs in vivo immunity in humans.

    PubMed

    Oliver, Samuel J; Macdonald, Jamie H; Harper Smith, Adam D; Lawley, Justin S; Gallagher, Carla A; Di Felice, Umberto; Walsh, Neil P

    2013-06-01

    The aim was to assess the effect of high altitude on the development of new immune memory (induction) using a contact sensitization model of in vivo immunity. We hypothesized that high-altitude exposure would impair induction of the in vivo immune response to a novel antigen, diphenylcyclopropenone (DPCP). DPCP was applied (sensitization) to the lower back of 27 rested controls at sea level and to ten rested mountaineers 28 hours after passive ascent to 3777 m. After sensitization, mountaineers avoided strenuous exercise for a further 24 hours, after which they completed alpine activities for 11-18 days. Exactly 4 weeks after sensitization, the strength of immune memory induction was quantified in rested mountaineers and controls at sea level, by measuring the response to a low, dose-series DPCP challenge, read at 48 hours as skin measures of edema (skinfold thickness) and redness (erythema). Compared with control responses, skinfold thickness and erythema were reduced in the mountaineers (skinfold thickness,-52%, p=0.01, d=0.86; erythema, -36%, p=0.02, d=0.77). These changes in skinfold thickness and erythema were related to arterial oxygen saturation (r=0.7, p=0.04), but not cortisol (r<0.1, p>0.79), at sensitization. In conclusion, this is the first study to show, using a contact sensitization model of in vivo immunity, that high altitude exposure impairs the development of new immunity in humans. PMID:23795734

  11. A Historically Significant Shield for In Vivo Measurements

    SciTech Connect

    Lynch, Timothy P.

    2007-08-01

    Due to the ubiquitous nature of ionizing radiation, in vivo measurement systems designed to measure low levels of radionuclides in people are usually enclosed within a high density shield. Lead, steel, earth, and water are just some of the materials that have been and are being used to shield the detectors from radiations of cosmic, atmospheric, and terrestrial origin. At many Department of Energy sites, the counting room shields are constructed of pre-world War II steel to reduce the background levels to achieve measurements with low minimum detectable activities (MDA). This is one example of what is commonly called low background steel in the in vivo industry vernacular. The name arises from the fact the steel was manufactured prior to the beginning of atmospheric testing of nuclear weapons in the 1940s. Consequently, the steel is not likely to be contaminated with fission or activation products from fallout. For high energy photons (600 keV In Vivo Radioassay and Research Facility (IVRRF) in Richland, Washington.

  12. In vivo dosimetry with silicon diodes in total body irradiation

    NASA Astrophysics Data System (ADS)

    Oliveira, F. F.; Amaral, L. L.; Costa, A. M.; Netto, T. G.

    2014-02-01

    The aim of this work is the characterization and application of silicon diode detectors for in vivo dosimetry in total body irradiation (TBI) treatments. It was evaluated the diode response with temperature, dose rate, gantry angulations and field size. A maximum response variation of 2.2% was obtained for temperature dependence. The response variation for dose rate and angular was within 1.2%. For field size dependence, the detector response increased with field until reach a saturation region, where no more primary radiation beam contributes for dose. The calibration was performed in a TBI setup. Different lateral thicknesses from one patient were simulated and then the calibration factors were determined by means of maximum depth dose readings. Subsequent to calibration, in vivo dosimetry measurements were performed. The response difference between diode readings and the prescribed dose for all treatments was below 4%. This difference is in agreement as recommended by the International Commission on Radiation Units and Measurements (ICRU), which is ±5%. The present work to test the applicability of a silicon diode dosimetry system for performing in vivo dose measurements in TBI techniques presented good results. These measurements demonstrated the value of diode dosimetry as a treatment verification method and its applicability as a part of a quality assurance program in TBI treatments.

  13. A historically significant shield for in vivo measurements.

    PubMed

    Lynch, Timothy P

    2007-08-01

    Due to the ubiquitous nature of ionizing radiation, in vivo measurement systems designed to measure low levels of radionuclides in people are usually enclosed within a high-density shield. Lead, steel, earth, and water are just some of the materials that have been and are being used to shield the detectors from radiations of cosmic, atmospheric, man-made, and terrestrial origin. At many Department of Energy sites, the counting room shields are constructed of pre-World War II steel to reduce the background levels in order to perform measurements that have low minimum detectable activities. The pre-World War II steel is commonly called low background steel in the in vivo industry vernacular. The low background descriptor comes from the fact the steel was manufactured prior to the beginning of atmospheric testing of nuclear weapons in the 1940's. Consequently, the steel is not likely to be contaminated with fission or activation products from fallout. For high energy photons (600 keV < E < 1500 keV), 30 cm of steel shielding significantly reduces the measured background radiation levels. This is the story of the unique steel that began as the hull of the U.S.S. Indiana and now forms a shielded room at the In Vivo Radiobioassay and Research Facility in Richland, Washington. PMID:17630635

  14. Structural imprints in vivo decode RNA regulatory mechanisms

    NASA Astrophysics Data System (ADS)

    Spitale, Robert C.; Flynn, Ryan A.; Zhang, Qiangfeng Cliff; Crisalli, Pete; Lee, Byron; Jung, Jong-Wha; Kuchelmeister, Hannes Y.; Batista, Pedro J.; Torre, Eduardo A.; Kool, Eric T.; Chang, Howard Y.

    2015-03-01

    Visualizing the physical basis for molecular behaviour inside living cells is a great challenge for biology. RNAs are central to biological regulation, and the ability of RNA to adopt specific structures intimately controls every step of the gene expression program. However, our understanding of physiological RNA structures is limited; current in vivo RNA structure profiles include only two of the four nucleotides that make up RNA. Here we present a novel biochemical approach, in vivo click selective 2'-hydroxyl acylation and profiling experiment (icSHAPE), which enables the first global view, to our knowledge, of RNA secondary structures in living cells for all four bases. icSHAPE of the mouse embryonic stem cell transcriptome versus purified RNA folded in vitro shows that the structural dynamics of RNA in the cellular environment distinguish different classes of RNAs and regulatory elements. Structural signatures at translational start sites and ribosome pause sites are conserved from in vitro conditions, suggesting that these RNA elements are programmed by sequence. In contrast, focal structural rearrangements in vivo reveal precise interfaces of RNA with RNA-binding proteins or RNA-modification sites that are consistent with atomic-resolution structural data. Such dynamic structural footprints enable accurate prediction of RNA-protein interactions and N6-methyladenosine (m6A) modification genome wide. These results open the door for structural genomics of RNA in living cells and reveal key physiological structures controlling gene expression.

  15. DEAD-box protein facilitated RNA folding in vivo

    PubMed Central

    Liebeg, Andreas; Mayer, Oliver

    2010-01-01

    In yeast mitochondria the DEAD-box helicase Mss116p is essential for respiratory growth by acting as group I and group II intron splicing factor. Here we provide the first structure-based insights into how Mss116p assists RNA folding in vivo. Employing an in vivo chemical probing technique, we mapped the structure of the ai5γ group II intron in different genetic backgrounds to characterize its intracellular fold. While the intron adopts the native conformation in the wt yeast strain, we found that the intron is able to form most of its secondary structure, but lacks its tertiary fold in the absence of Mss116p. This suggests that ai5γ is largely unfolded in the mss116-knockout strain and requires the protein at an early step of folding. Notably, in this unfolded state misfolded substructures have not been observed. As most of the protein-induced conformational changes are located within domain D1, Mss116p appears to facilitate the formation of this largest domain, which is the scaffold for docking of other intron domains. These findings suggest that Mss116p assists the ordered assembly of the ai5γ intron in vivo. PMID:21045551

  16. In vivo enzyme activity in inborn errors of metabolism

    SciTech Connect

    Thompson, G.N.; Walter, J.H.; Leonard, J.V.; Halliday, D. )

    1990-08-01

    Low-dose continuous infusions of (2H5)phenylalanine, (1-13C)propionate, and (1-13C)leucine were used to quantitate phenylalanine hydroxylation in phenylketonuria (PKU, four subjects), propionate oxidation in methylmalonic acidaemia (MMA, four subjects), and propionic acidaemia (PA, four subjects) and leucine oxidation in maple syrup urine disease (MSUD, four subjects). In vivo enzyme activity in PKU, MMA, and PA subjects was similar to or in excess of that in adult controls (range of phenylalanine hydroxylation in PKU, 3.7 to 6.5 mumol/kg/h, control 3.2 to 7.9, n = 7; propionate oxidation in MMA, 15.2 to 64.8 mumol/kg/h, and in PA, 11.1 to 36.0, control 5.1 to 19.0, n = 5). By contrast, in vivo leucine oxidation was undetectable in three of the four MSUD subjects (less than 0.5 mumol/kg/h) and negligible in the remaining subject (2 mumol/kg/h, control 10.4 to 15.7, n = 6). These results suggest that significant substrate removal can be achieved in some inborn metabolic errors either through stimulation of residual enzyme activity in defective enzyme systems or by activation of alternate metabolic pathways. Both possibilities almost certainly depend on gross elevation of substrate concentrations. By contrast, only minimal in vivo oxidation of leucine appears possible in MSUD.

  17. Histone acetyltransferase inhibitors block neuroblastoma cell growth in vivo

    PubMed Central

    Gajer, J M; Furdas, S D; Gründer, A; Gothwal, M; Heinicke, U; Keller, K; Colland, F; Fulda, S; Pahl, H L; Fichtner, I; Sippl, W; Jung, M

    2015-01-01

    We have previously described novel histone acetyltransferase (HAT) inhibitors that block neuroblastoma cell growth in vitro. Here we show that two selected pyridoisothiazolone HAT inhibitors, PU139 and PU141, induce cellular histone hypoacetylation and inhibit growth of several neoplastic cell lines originating from different tissues. Broader in vitro selectivity profiling shows that PU139 blocks the HATs Gcn5, p300/CBP-associated factor (PCAF), CREB (cAMP response element-binding) protein (CBP) and p300, whereas PU141 is selective toward CBP and p300. The pan-inhibitor PU139 triggers caspase-independent cell death in cell culture. Both inhibitors block growth of SK-N-SH neuroblastoma xenografts in mice and the PU139 was shown to synergize with doxorubicin in vivo. The latter also reduces histone lysine acetylation in vivo at concentrations that block neoplastic xenograft growth. This is one of the very few reports on hypoacetylating agents with in vivo anticancer activity. PMID:25664930

  18. Immunotherapy of Malignancy by in vivo Gene Transfer into Tumors

    NASA Astrophysics Data System (ADS)

    Plautz, Gregory E.; Yang, Zhi-Yong; Wu, Bei-Yue; Gao, Xiang; Huang, Leaf; Nabel, Gary J.

    1993-05-01

    The immune system confers protection against a variety of pathogens and contributes to the surveillance and destruction of neoplastic cells. Several cell types participate in the recognition and lysis of tumors, and appropriate immune stimulation provides therapeutic effects in malignancy. Foreign major histocompatibility complex (MHC) proteins also serve as a potent stimulus to the immune system. In this report, a foreign MHC gene was introduced directly into malignant tumors in vivo in an effort to stimulate tumor rejection. In contrast to previous attempts to induce tumor immunity by cell-mediated gene transfer, the recombinant gene was introduced directly into tumors in vivo. Expression of the murine class I H-2K^s gene within the CT26 mouse colon adenocarcinoma (H-2K^d) or the MCA 106 fibrosarcoma (H-2K^b) induced a cytotoxic T-cell response to H-2K^s and, more importantly, to other antigens present on unmodified tumor cells. This immune response attenuated tumor growth and caused complete tumor regression in many cases. Direct gene transfer in vivo can therefore induce cell-mediated immunity against specific gene products, which provides an immunotherapeutic effect for malignancy, and potentially can be applied to the treatment of cancer and infectious diseases in man.

  19. Favipiravir elicits antiviral mutagenesis during virus replication in vivo

    PubMed Central

    Arias, Armando; Thorne, Lucy; Goodfellow, Ian

    2014-01-01

    Lethal mutagenesis has emerged as a novel potential therapeutic approach to treat viral infections. Several studies have demonstrated that increases in the high mutation rates inherent to RNA viruses lead to viral extinction in cell culture, but evidence during infections in vivo is limited. In this study, we show that the broad-range antiviral nucleoside favipiravir reduces viral load in vivo by exerting antiviral mutagenesis in a mouse model for norovirus infection. Increased mutation frequencies were observed in samples from treated mice and were accompanied with lower or in some cases undetectable levels of infectious virus in faeces and tissues. Viral RNA isolated from treated animals showed reduced infectivity, a feature of populations approaching extinction during antiviral mutagenesis. These results suggest that favipiravir can induce norovirus mutagenesis in vivo, which in some cases leads to virus extinction, providing a proof-of-principle for the use of favipiravir derivatives or mutagenic nucleosides in the clinical treatment of noroviruses. DOI: http://dx.doi.org/10.7554/eLife.03679.001 PMID:25333492

  20. A guide to studying human hair follicle cycling in vivo

    PubMed Central

    Oh, Ji Won; Kloepper, Jennifer; Langan, Ewan A.; Kim, Yongsoo; Yeo, Joongyeub; Kim, Min Ji; Hsi, Tsai-Ching; Rose, Christian; Yoon, Ghil Suk; Lee, Seok-Jong; Seykora, John; Kim, Jung Chul; Sung, Young Kwan

    2015-01-01

    Hair follicles (HFs) undergo life-long cyclical transformations, progressing through stages of rapid growth (anagen), regression (catagen), and relative “quiescence” (telogen). Since HF cycling abnormalities underlie many human hair growth disorders, the accurate classification of individual cycle stages within skin biopsies is clinically important and essential for hair research. For preclinical human hair research purposes, human scalp skin can be xenografted onto immunocompromised mice to study human HF cycling and manipulate long-lasting anagen in vivo. While available for mice, a comprehensive guide on how to recognize different human hair cycle stages in vivo is lacking. Here, we present such a guide, which uses objective, well-defined, and reproducible criteria and integrates simple morphological indicators with advanced, (immuno)-histochemical markers. This guide also characterizes human HF cycling in xenografts and highlights the utility of this model for in vivo hair research. Detailed schematic drawings and representative micrographs provide examples of how best to identify human HF stages, even in sub-optimally sectioned tissue, and practical recommendations are given for designing human-on-mouse hair cycle experiments. Thus, this guide seeks to offer a benchmark for human hair cycle stage classification, for both hair research experts and newcomers to the field. PMID:26763421

  1. Portable XRF Technology to Quantify Pb in Bone In Vivo.

    PubMed

    Specht, Aaron James; Weisskopf, Marc; Nie, Linda Huiling

    2014-01-01

    Lead is a ubiquitous toxicant. Bone lead has been established as an important biomarker for cumulative lead exposures and has been correlated with adverse health effects on many systems in the body. K-shell X-ray fluorescence (KXRF) is the standard method for measuring bone lead, but this approach has many difficulties that have limited the widespread use of this exposure assessment method. With recent advancements in X-ray fluorescence (XRF) technology, we have developed a portable system that can quantify lead in bone in vivo within 3 minutes. Our study investigated improvements to the system, four calibration methods, and system validation for in vivo measurements. Our main results show that the detection limit of the system is 2.9 ppm with 2 mm soft tissue thickness, the best calibration method for in vivo measurement is background subtraction, and there is strong correlation between KXRF and portable LXRF bone lead results. Our results indicate that the technology is ready to be used in large human population studies to investigate adverse health effects of lead exposure. The portability of the system and fast measurement time should allow for this technology to greatly advance the research on lead exposure and public/environmental health. PMID:26317033

  2. Portable XRF Technology to Quantify Pb in Bone In Vivo

    PubMed Central

    Specht, Aaron James; Weisskopf, Marc; Nie, Linda Huiling

    2014-01-01

    Lead is a ubiquitous toxicant. Bone lead has been established as an important biomarker for cumulative lead exposures and has been correlated with adverse health effects on many systems in the body. K-shell X-ray fluorescence (KXRF) is the standard method for measuring bone lead, but this approach has many difficulties that have limited the widespread use of this exposure assessment method. With recent advancements in X-ray fluorescence (XRF) technology, we have developed a portable system that can quantify lead in bone in vivo within 3 minutes. Our study investigated improvements to the system, four calibration methods, and system validation for in vivo measurements. Our main results show that the detection limit of the system is 2.9 ppm with 2 mm soft tissue thickness, the best calibration method for in vivo measurement is background subtraction, and there is strong correlation between KXRF and portable LXRF bone lead results. Our results indicate that the technology is ready to be used in large human population studies to investigate adverse health effects of lead exposure. The portability of the system and fast measurement time should allow for this technology to greatly advance the research on lead exposure and public/environmental health. PMID:26317033

  3. Simultaneous in vivo positron emission tomography and magnetic resonance imaging

    PubMed Central

    Catana, Ciprian; Procissi, Daniel; Wu, Yibao; Judenhofer, Martin S.; Qi, Jinyi; Pichler, Bernd J.; Jacobs, Russell E.; Cherry, Simon R.

    2008-01-01

    Positron emission tomography (PET) and magnetic resonance imaging (MRI) are widely used in vivo imaging technologies with both clinical and biomedical research applications. The strengths of MRI include high-resolution, high-contrast morphologic imaging of soft tissues; the ability to image physiologic parameters such as diffusion and changes in oxygenation level resulting from neuronal stimulation; and the measurement of metabolites using chemical shift imaging. PET images the distribution of biologically targeted radiotracers with high sensitivity, but images generally lack anatomic context and are of lower spatial resolution. Integration of these technologies permits the acquisition of temporally correlated data showing the distribution of PET radiotracers and MRI contrast agents or MR-detectable metabolites, with registration to the underlying anatomy. An MRI-compatible PET scanner has been built for biomedical research applications that allows data from both modalities to be acquired simultaneously. Experiments demonstrate no effect of the MRI system on the spatial resolution of the PET system and <10% reduction in the fraction of radioactive decay events detected by the PET scanner inside the MRI. The signal-to-noise ratio and uniformity of the MR images, with the exception of one particular pulse sequence, were little affected by the presence of the PET scanner. In vivo simultaneous PET and MRI studies were performed in mice. Proof-of-principle in vivo MR spectroscopy and functional MRI experiments were also demonstrated with the combined scanner. PMID:18319342

  4. Aspartame induces angiogenesis in vitro and in vivo models.

    PubMed

    Yesildal, F; Aydin, F N; Deveci, S; Tekin, S; Aydin, I; Mammadov, R; Fermanli, O; Avcu, F; Acikel, C H; Ozgurtas, T

    2015-03-01

    Angiogenesis is the process of generating new blood vessels from preexisting vessels and is considered essential in many pathological conditions. The purpose of the present study is to evaluate the effect of aspartame on angiogenesis in vivo chick chorioallantoic membrane (CAM) and wound-healing models as well as in vitro 2,3-bis-2H-tetrazolium-5-carboxanilide (XTT) and tube formation assays. In CAM assay, aspartame increased angiogenesis in a concentration-dependent manner. Compared with the control group, aspartame has significantly increased vessel proliferation (p < 0.001). In addition, in vivo rat model of skin wound-healing study showed that aspartame group had better healing than control group, and this was statistically significant at p < 0.05. There was a slight proliferative effect of aspartame on human umbilical vein endothelial cells on XTT assay in vitro, but it was not statistically significant; and there was no antiangiogenic effect of aspartame on tube formation assay in vitro. These results provide evidence that aspartame induces angiogenesis in vitro and in vivo; so regular use may have undesirable effect on susceptible cases. PMID:24925367

  5. Experimental aspects of in vitro and in vivo photochemotherapy.

    PubMed

    Patrice, T; Praloran, V; Le Bodic, M F; Le Bodic, L

    1986-06-01

    The selectivity of in vitro photodynamic reactions and the in vivo effects induced by PRT, whether the irradiation is applied interstitially or externally, still remains unclear. In vitro studies were performed using leukemic cell lines and syngeneic normal hemopoietic progenitors. For these, cells incubated with hematoporphyrin derivative (HPD) and non-incubated cells were irradiated with an argon laser. Data were obtained as the count of cell colonies found after a 7-day incubation period on semi-solid collagen gel medium. In vivo studies employed the HT 29 tumor model grafted into nude mice. Both animals injected with HPD and non-infected controls were irradiated with a dye laser pumped by an argon laser (Coherent) using a 400 micron optic fiber located either at a distance of 65 mm from the skin or inserted into the tumor. The temperature increase occurring during PRT was measured using non-absorbing thermocouples. In vitro, after HPD treatment and argon irradiation leukemic cells showed a greater phototoxicity (greater than 2 log10) than did the normal cells (0.25 log10). In vivo, when the heat rise is very similar (less than 4 degrees C) in both the tissues irradiated externally and those irradiated interstitially after HPD injection, histological examination of these did not reveal any quantitative differences (90% of tumor mass). These results are discussed. PMID:2944549

  6. Resurrection of DNA function in vivo from an extinct genome.

    PubMed

    Pask, Andrew J; Behringer, Richard R; Renfree, Marilyn B

    2008-01-01

    There is a burgeoning repository of information available from ancient DNA that can be used to understand how genomes have evolved and to determine the genetic features that defined a particular species. To assess the functional consequences of changes to a genome, a variety of methods are needed to examine extinct DNA function. We isolated a transcriptional enhancer element from the genome of an extinct marsupial, the Tasmanian tiger (Thylacinus cynocephalus or thylacine), obtained from 100 year-old ethanol-fixed tissues from museum collections. We then examined the function of the enhancer in vivo. Using a transgenic approach, it was possible to resurrect DNA function in transgenic mice. The results demonstrate that the thylacine Col2A1 enhancer directed chondrocyte-specific expression in this extinct mammalian species in the same way as its orthologue does in mice. While other studies have examined extinct coding DNA function in vitro, this is the first example of the restoration of extinct non-coding DNA and examination of its function in vivo. Our method using transgenesis can be used to explore the function of regulatory and protein-coding sequences obtained from any extinct species in an in vivo model system, providing important insights into gene evolution and diversity. PMID:18493600

  7. Resurrection of DNA Function In Vivo from an Extinct Genome

    PubMed Central

    Pask, Andrew J.; Behringer, Richard R.; Renfree, Marilyn B.

    2008-01-01

    There is a burgeoning repository of information available from ancient DNA that can be used to understand how genomes have evolved and to determine the genetic features that defined a particular species. To assess the functional consequences of changes to a genome, a variety of methods are needed to examine extinct DNA function. We isolated a transcriptional enhancer element from the genome of an extinct marsupial, the Tasmanian tiger (Thylacinus cynocephalus or thylacine), obtained from 100 year-old ethanol-fixed tissues from museum collections. We then examined the function of the enhancer in vivo. Using a transgenic approach, it was possible to resurrect DNA function in transgenic mice. The results demonstrate that the thylacine Col2A1 enhancer directed chondrocyte-specific expression in this extinct mammalian species in the same way as its orthologue does in mice. While other studies have examined extinct coding DNA function in vitro, this is the first example of the restoration of extinct non-coding DNA and examination of its function in vivo. Our method using transgenesis can be used to explore the function of regulatory and protein-coding sequences obtained from any extinct species in an in vivo model system, providing important insights into gene evolution and diversity. PMID:18493600

  8. 2D luminescence imaging of pH in vivo

    PubMed Central

    Schreml, Stephan; Meier, Robert J.; Wolfbeis, Otto S.; Landthaler, Michael; Szeimies, Rolf-Markus; Babilas, Philipp

    2011-01-01

    Luminescence imaging of biological parameters is an emerging field in biomedical sciences. Tools to study 2D pH distribution are needed to gain new insights into complex disease processes, such as wound healing and tumor metabolism. In recent years, luminescence-based methods for pH measurement have been developed. However, for in vivo applications, especially for studies on humans, biocompatibility and reliability under varying conditions have to be ensured. Here, we present a referenced luminescent sensor for 2D high-resolution imaging of pH in vivo. The ratiometric sensing scheme is based on time-domain luminescence imaging of FITC and ruthenium(II)tris-(4,7-diphenyl-1,10-phenanthroline). To create a biocompatible 2D sensor, these dyes were bound to or incorporated into microparticles (aminocellulose and polyacrylonitrile), and particles were immobilized in polyurethane hydrogel on transparent foils. We show sensor precision and validity by conducting in vitro and in vivo experiments, and we show the versatility in imaging pH during physiological and chronic cutaneous wound healing in humans. Implementation of this technique may open vistas in wound healing, tumor biology, and other biomedical fields. PMID:21262842

  9. Synchronization among neuronal pools without common inputs: in vivo study.

    PubMed

    Brama, Haya; Guberman, Shoshana; Abeles, Moshe; Stern, Edward; Kanter, Ido

    2015-11-01

    Periodic synchronization of activity among neuronal pools has been related to substantial neural processes and information throughput in the neocortical network. However, the mechanisms of generating such periodic synchronization among distributed pools of neurons remain unclear. We hypothesize that to a large extent there is interplay between the topology of the neocortical networks and their reverberating modes of activity. The firing synchronization is governed by a nonlocal mechanism, the network delay loops, such that distant neuronal pools without common drives can be synchronized. This theoretical interplay between network topology and the synchronized mode is verified using an iterative procedure of a single intracellularly recorded neuron in vivo, imitating the dynamics of the entire network. The input is injected to the neuron via the recording electrode as current and computed from the filtered, evoked spikes of its pre-synaptic sources, previously emulated by the same neuron. In this manner we approximate subthreshold synaptic inputs from afferent neuronal pools to the neuron. Embedding the activity of these recurrent motifs in the intact brain allows us to measure the effects of connection probability, synaptic strength, and ongoing activity on the neuronal synchrony. Our in vivo experiments indicate that an initial stimulus given to a single pool is dynamically evolving into the formations of zero-lag and cluster synchronization. By applying results from theoretical models and in vitro experiments to in vivo activity in the intact brain, we support the notion that this mechanism may account for the binding activity across distributed brain areas. PMID:25230822

  10. In vivo imaging with near-infrared fluorescence lifetime contrast

    NASA Astrophysics Data System (ADS)

    Akers, Walter J.; Berezin, Mikhail Y.; Lee, Hyeran; Achilefu, Samuel

    2009-02-01

    Fluorescence imaging is a mainstay of biomedical research, allowing detection of molecular events in both fixed and living cells, tissues and whole animals. Such high resolution fluorescence imaging is hampered by unwanted signal from intrinsic background fluorescence and scattered light. The signal to background ratio can be improved by using extrinsic contrast agents and greatly enhanced by multispectral imaging methods. Unfortunately, these methods are insufficient for deep tissue imaging where high contrast and speedy acquisition are necessary. Fluorescence lifetime (FLT) is an inherent characteristic of each fluorescent species that can be independent of intensity and spectral properties. Accordingly, FLT-based detection provides an additional contrast mechanism to optical measurements. This contrast is particularly important in the near-infrared (NIR) due to relative transparency of tissue as well as the broad absorption and emission spectra of dyes that are active in this region. Here we report comparative analysis of signal distribution of several NIR fluorescent polymethine dyes in living mice and their correlations with lifetimes obtained in vitro using solution models. The FLT data obtained from dyes dissolved in serum albumin solution correlated well with FLTs measured in vivo. Thus the albumin solution model could be used as a good predictive model for in vivo FLT behavior of newly developed fluorescent reporters. Subsequent experiments in vivo, including monitoring slow release kinetics and detecting proteinuria, demonstrate the complementary nature of FLT for fluorescence intensity imaging.

  11. A Guide to Studying Human Hair Follicle Cycling In Vivo.

    PubMed

    Oh, Ji Won; Kloepper, Jennifer; Langan, Ewan A; Kim, Yongsoo; Yeo, Joongyeub; Kim, Min Ji; Hsi, Tsai-Ching; Rose, Christian; Yoon, Ghil Suk; Lee, Seok-Jong; Seykora, John; Kim, Jung Chul; Sung, Young Kwan; Kim, Moonkyu; Paus, Ralf; Plikus, Maksim V

    2016-01-01

    Hair follicles (HFs) undergo lifelong cyclical transformations, progressing through stages of rapid growth (anagen), regression (catagen), and relative "quiescence" (telogen). Given that HF cycling abnormalities underlie many human hair growth disorders, the accurate classification of individual cycle stages within skin biopsies is clinically important and essential for hair research. For preclinical human hair research purposes, human scalp skin can be xenografted onto immunocompromised mice to study human HF cycling and manipulate long-lasting anagen in vivo. Although available for mice, a comprehensive guide on how to recognize different human hair cycle stages in vivo is lacking. In this article, we present such a guide, which uses objective, well-defined, and reproducible criteria, and integrates simple morphological indicators with advanced, (immuno)-histochemical markers. This guide also characterizes human HF cycling in xenografts and highlights the utility of this model for in vivo hair research. Detailed schematic drawings and representative micrographs provide examples of how best to identify human HF stages, even in suboptimally sectioned tissue, and practical recommendations are given for designing human-on-mouse hair cycle experiments. Thus, this guide seeks to offer a benchmark for human hair cycle stage classification, for both hair research experts and newcomers to the field. PMID:26763421

  12. Identification of Enterococcus faecalis antigens specifically expressed in vivo

    PubMed Central

    Shet, Uttom K.; Park, Sang-Won; Lim, Hyun-Pil; Yun, Kwi-Dug; Kang, Seong Soo; Kim, Se Eun

    2015-01-01

    Objectives Molecular mechanism of the pathogenicity of Enterococcus faecalis (E. faecalis), a suspected endodontic pathogen, has not yet been adequately elucidated due to limited information on its virulence factors. Here we report the identification of in vivo expressed antigens of E. faecalis by using a novel immunoscreening technique called change-mediated antigen technology (CMAT) and an experimental animal model of endodontic infection. Materials and Methods Among 4,500 E. coli recombinant clones screened, 19 positive clones reacted reproducibly with hyperimmune sera obtained from rabbits immunized with E. faecalis cells isolated from an experimental endodontic infection. DNA sequences from 16 of these in vivo-induced (IVI) genes were determined. Results Identified protein antigens of E. faecalis included enzymes involved in housekeeping functions, copper resistance protein, putative outer membrane proteins, and proteins of unknown function. Conclusions In vivo expressed antigens of E. faecalis could be identified by using a novel immune-screening technique CMAT and an experimental animal model of endodontic infection. Detailed analysis of these IVI genes will lead to a better understanding of the molecular mechanisms involved in the endodontic infection of E. faecalis. PMID:26587417

  13. Organelle acidification negatively regulates vacuole membrane fusion in vivo.

    PubMed

    Desfougères, Yann; Vavassori, Stefano; Rompf, Maria; Gerasimaite, Ruta; Mayer, Andreas

    2016-01-01

    The V-ATPase is a proton pump consisting of a membrane-integral V0 sector and a peripheral V1 sector, which carries the ATPase activity. In vitro studies of yeast vacuole fusion and evidence from worms, flies, zebrafish and mice suggested that V0 interacts with the SNARE machinery for membrane fusion, that it promotes the induction of hemifusion and that this activity requires physical presence of V0 rather than its proton pump activity. A recent in vivo study in yeast has challenged these interpretations, concluding that fusion required solely lumenal acidification but not the V0 sector itself. Here, we identify the reasons for this discrepancy and reconcile it. We find that acute pharmacological or physiological inhibition of V-ATPase pump activity de-acidifies the vacuole lumen in living yeast cells within minutes. Time-lapse microscopy revealed that de-acidification induces vacuole fusion rather than inhibiting it. Cells expressing mutated V0 subunits that maintain vacuolar acidity were blocked in this fusion. Thus, proton pump activity of the V-ATPase negatively regulates vacuole fusion in vivo. Vacuole fusion in vivo does, however, require physical presence of a fusion-competent V0 sector. PMID:27363625

  14. Effect of desipramine on dopamine receptor binding in vivo

    SciTech Connect

    Suhara, Tetsuya Jikei Univ., Tokyo ); Inoue, Osamu; Kobayasi, Kaoru )

    1990-01-01

    Effect of desipramine on the in vivo binding of {sup 3}H-SCH23390 and {sup 3}H-N-methylspiperone ({sup 3}H-NMSP) in mouse striatum was studied. The ratio of radioactivity in the striatum to that in the cerebellum at 15 min after i.v. injection of {sup 3}H-SCH23390 or 45 min after injection of {sup 3}H-NMSP were used as indices of dopamine D1 or D2 receptor binding in vivo, respectively. In vivo binding of D1 and D2 receptors was decreased in a dose-dependent manner by acute treatment with desipramine (DMI). A saturation experiment suggested that the DMI-induced reduction in the binding was mainly due to the decrease in the affinity of both receptors. No direct interactions between the dopamine receptors and DMI were observed in vitro by the addition of 1 mM of DMI into striatal homogenate. Other antidepressants such as imipramine, clomipramine, maprotiline and mianserin also decreased the binding of dopamine D1 and D2 receptors. The results indicated an important role of dopamine receptors in the pharmacological effect of antidepressants.

  15. mito-QC illuminates mitophagy and mitochondrial architecture in vivo.

    PubMed

    McWilliams, Thomas G; Prescott, Alan R; Allen, George F G; Tamjar, Jevgenia; Munson, Michael J; Thomson, Calum; Muqit, Miratul M K; Ganley, Ian G

    2016-08-01

    Autophagic turnover of mitochondria, termed mitophagy, is proposed to be an essential quality-control (QC) mechanism of pathophysiological relevance in mammals. However, if and how mitophagy proceeds within specific cellular subtypes in vivo remains unclear, largely because of a lack of tractable tools and models. To address this, we have developed "mito-QC," a transgenic mouse with a pH-sensitive fluorescent mitochondrial signal. This allows the assessment of mitophagy and mitochondrial architecture in vivo. Using confocal microscopy, we demonstrate that mito-QC is compatible with classical and contemporary techniques in histochemistry and allows unambiguous in vivo detection of mitophagy and mitochondrial morphology at single-cell resolution within multiple organ systems. Strikingly, our model uncovers highly enriched and differential zones of mitophagy in the developing heart and within specific cells of the adult kidney. mito-QC is an experimentally advantageous tool of broad relevance to cell biology researchers within both discovery-based and translational research communities. PMID:27458135

  16. Circulation times of cancer cells by in vivo flow cytometry

    NASA Astrophysics Data System (ADS)

    Zhang, Li; Li, Yan; Gu, Zhengqin; Chen, Tong; Wang, Cheng; Wei, Xunbin

    2012-03-01

    Liver cancer is one of the most common malignancies in the world, with approximately 1,000,000 cases reported every year. Hepatocellular carcinoma may metastasize to lung, bones, kidney, and many other organs. Surgical resection, liver transplantation, chemotherapy and radiation therapy are the foundation of current HCC therapies. However the outcomes are poor: the survival rate is almost zero for metastatic HCC patients. Molecular mechanisms of HCC metastasis need to be understood better and new therapies must be developed. A recently developed "in vivo flow cytometer" combined with real-time confocal fluorescence imaging are used to assess spreading and the circulation kinetics of liver tumor cells. The in vivo flow cytometer has the capability to detect and quantify continuously the number and flow characteristics of fluorescently labeled cells in vivo in real time without extracting blood sample. We have measured the depletion kinetics of two related human HCC cell lines, high-metastatic HCCLM3 cells and low-metastatic HepG2 cells, which were from the same origin and obtained by repetitive screenings in mice. >60% HCCLM3 cells are depleted within the first hour. Interestingly, the low-metastatic HepG2 cells possess noticeably slower depletion kinetics. In comparison, <40% HepG2 cells are depleted within the first hour. The differences in depletion kinetics might provide insights into early metastasis processes.

  17. Cellular metabolites modulate in vivo signaling of Arabidopsis cryptochrome-1

    PubMed Central

    El-Esawi, Mohamed; Glascoe, Austin; Engle, Dorothy; Ritz, Thorsten; Link, Justin; Ahmad, Margaret

    2015-01-01

    Cryptochromes are blue-light absorbing flavoproteins with multiple signaling roles. In plants, cryptochrome (cry1, cry2) biological activity has been linked to flavin photoreduction via an electron transport chain to the protein surface comprising 3 evolutionarily conserved tryptophan residues known as the ‘Trp triad.’ Mutation of any of the Trp triad residues abolishes photoreduction in isolated cryptochrome protein in vitro and therefore had been suggested as essential for electron transfer to the flavin. However, photoreduction of the flavin in Arabidopsis cry2 proteins occurs in vivo even with mutations in the Trp triad, indicating the existence of alternative electron transfer pathways to the flavin. These pathways are potentiated by metabolites in the intracellular environment including ATP, ADP, AMP, and NADH. In the present work we extend these observations to Arabidopsis cryptochrome 1 and demonstrate that Trp triad substitution mutants at W400F and W324F positions which are not photoreduced in vitro can be photoreduced in whole cell extracts, albeit with reduced efficiency. We further show that the flavin signaling state (FADH°) is stabilized in an in vivo context. These data illustrate that in vivo modulation by metabolites in the cellular environment may play an important role in cryptochrome signaling, and are discussed with respect to possible effects on the conformation of the C-terminal domain to generate the biologically active conformational state. PMID:26313597

  18. Caspase 3 in dying tumor cells mediates post-irradiation angiogenesis

    PubMed Central

    Zhang, Zhengxiang; Yu, Yang; Cheng, Jin; Gong, Yanping; Li, Chuan-Yuan; Huang, Qian

    2015-01-01

    Cytotoxic radiotherapy unfavorably induces tumor cells to generate various proangiogenic substances, promoting post-irradiation angiogenesis (PIA), which is one of major causes of radiotherapy failure. Though several studies have reported some mechanisms behind PIA, they have not yet described the beginning proangiogenic motivator buried in the irradiated microenvironment. In this work, we revealed that dying tumor cells induced by irradiation prompted PIA via a caspase 3 dependent mechanism. Proteolytic inactivation of caspase 3 in dying tumor cells by transducing a dominant-negative version weakened proangiogenic effects in vitro and in vivo. In addition, inhibition of caspase 3 activity suppressed tumor angiogenesis and tumorigenesis in xenograft mouse model. Importantly, we identified vascular endothelial growth factor (VEGF)-A as a downstream proangiogenic factor regulated by caspase 3 possibly through Akt signaling. Collectively, these findings indicated that besides acting as a key executioner in apoptosis, caspase 3 in dying tumor cells may play a central role in driving proangiogenic response after irradiation. Thus, radiotherapy in combination with caspase 3 inhibitors may be a novel promising therapeutic strategy to reduce tumor recurrence due to restrained PIA. PMID:26431328

  19. Rapid prototyping of extrusion dies using layer-based techniques

    SciTech Connect

    Misiolek, W.Z.; Winther, K.T.; Prats, A.E.; Rock, S.J.

    1999-02-01

    Extrusion die design and development often requires significant craftsman skill and iterative improvement to arrive at a production-ready die geometry. Constructing the dies used during this iterative process from layers, rather than from one solid block of material, offers unique opportunities to improve die development efficiency when coupled with concepts drawn from the rapid prototyping field. This article presents a proof-of-concept illustrating the potential utility of layer-based extrusion dies for the die design and fabrication process. The major benefits include greater flexibility in the design process, a more efficient, automated fabrication technique, and a means for performing localized die modifications and repairs.

  20. EPID based in vivo dosimetry system: clinical experience and results.

    PubMed

    Celi, Sofia; Costa, Emilie; Wessels, Claas; Mazal, Alejandro; Fourquet, Alain; Francois, Pascal

    2016-01-01

    Mandatory in several countries, in vivo dosimetry has been recognized as one of the next milestones in radiation oncology. Our department has implemented clinically an EPID based in vivo dosimetry system, EPIgray, by DOSISOFT S.A., since 2006. An analysis of the measurements per linac and energy over a two-year period was performed, which included a more detailed examination per technique and treat-ment site over a six-month period. A comparison of the treatment planning system doses and the doses estimated by EPIgray shows a mean of the differences of 1.9% (± 5.2%) for the two-year period. The 3D conformal treatment plans had a mean dose difference of 2.0% (± 4.9%), while for intensity-modulated radiotherapy and volumetric-modulated arc therapy treatments the mean dose difference was -3.0 (± 5.3%) and -2.5 (± 5.2%), respectively. In addition, root cause analyses were conducted on the in vivo dosimetry measurements of two breast cancer treatment techniques, as well as prostate treatments with intensity-modulated radiotherapy and volumetric-modulated arc therapy. During the breast study, the dose differences of breast treatments in supine position were correlated to patient setup and EPID positioning errors. Based on these observations, an automatic image shift correc-tion algorithm is developed by DOSIsoft S.A. The prostate study revealed that beams and arcs with out-of-tolerance in vivo dosimetry results tend to have more complex modulation and a lower exposure of the points of interest. The statistical studies indicate that in vivo dosimetry with EPIgray has been successfully imple-mented for classical and complex techniques in clinical routine at our institution. The additional breast and prostate studies exhibit the prospects of EPIgray as an easy supplementary quality assurance tool. The validation, the automatization, and the reduction of false-positive results represent an important step toward adaptive radiotherapy with EPIgray. PMID:27167283

  1. Fiber optic confocal microscope: In vivo precancer detection

    NASA Astrophysics Data System (ADS)

    Carlson, Kristen Dawn

    Cancer is a significant public health problem worldwide. Many cancers originate as precancerous lesions in the epithelium which, when removed in sufficient time, can prevent progression to cancer. However, current detection techniques are typically time-consuming and expensive, limiting their acceptance and accessibility. Optical techniques, such as confocal microscopy, have significant potential to provide clinicians with real-time, high-resolution images of cells and tissue without tissue removal. These images of cell morphology and tissue architecture can be used to characterize tissue and determine the presence or extent of precancer and cancer. This dissertation explores the instrumentation and application of fiber optic reflectance confocal microscopy for in vivo precancer detection. The first part of the dissertation presents in vivo imaging of suspicious lesions in the human uterine cervix and oral mucosa using a fiber bundle based confocal microscope with a complex glass miniature objective lens. Images are analyzed quantitatively and qualitatively to determine the potential of this technology in vivo. An analysis of nuclear density from images of 30 cervical epithelium sites shows differentiation between normal and precancerous sites. Similarly, images from 20 oral mucosa sites demonstrate changes in nuclear density and tissue architecture indicative of progression of precancer and cancer. In addition to this multi-fiber confocal microscope used with a glass objective lens for the clinical studies, imaging of tissue samples has been accomplished with the same confocal system using an injection molded plastic miniature objective lens demonstrating comparable optical quality for a significantly less expensive optical component. Finally, a benchtop prototype of a single fiber confocal microscope using a gimbaled two-axis MEMS scanner has been designed and constructed. Imaging of a resolution target and cellular samples demonstrates sufficient resolution and

  2. Why do patients with lupus nephritis die?

    PubMed Central

    Correia, P; Cameron, J S; Lian, J D; Hicks, J; Ogg, C S; Williams, D G; Chantler, C; Haycock, D G

    1985-01-01

    Over 20 years 42 of 138 patients with systemic lupus erythematosus "died"--that is, suffered actual death or went into terminal renal failure, or both; data from 41 were available for analysis. In most patients the causes of death were multiple. Twenty seven patients went into terminal renal failure, of whom 25 were offered dialysis treatment. Three regained renal function later, 12 survived on dialysis or with functioning kidney allografts--almost all with inactive lupus--but 13 died after starting dialysis, most within a few weeks or months. The principal causes were active lupus or infection. In those patients with renal failure after rapid deterioration in renal function (n = 14) there were nine deaths, while of 10 patients with a slow evolution into renal failure, only four died. Four patients with impaired and 10 with normal renal function died, again most often from complications of lupus or from infection. Vascular disease was a major cause of death in seven patients, all but two of whom were young; of 15 postmortem examinations, eight showed severe coronary artery atheroma, and three surviving patients required coronary bypass operations. Analysis of the timing of death or entry into renal failure showed that in 12 out of 13 patients who died within two years of onset the lupus was judged to be active, while this was true in only eight out of 19 patients who died later. Six of the seven vascular deaths occurred later than two years from onset, while only nine of 26 renal "deaths" occurred before two years; deaths from infections (n = 13) were distributed equally. Despite this and aggressive treatment of active disease, the principal cause of actual death was uncontrolled lupus. PMID:3917713

  3. Die Europäische Union, die Europäische Gemeinschaft und ihre Rechtsordnung, die Europäische Lebensmittelkontrolle

    NASA Astrophysics Data System (ADS)

    Gallhoff, Gudrun; Rimkus, Gerhard G.

    Die Europäische Union (EU) ist ein Zusammenschluss von siebenundzwanzig unabhängigen Staaten, um deren wirtschaftliche, politische und soziale Zusammenarbeit zu verstärken. Seit 1. Mai 2007 hat sie die folgenden Mitglieder: Österreich, Belgien, Bulgarien, Dänemark, Finnland, Frankreich, Deutschland, Griechenland, Irland, Italien, Luxemburg, die Niederlande, Portugal, Spanien, Schweden, das Vereinigte Königreich von Großbritannien und Nordirland, Zypern, die Tschechische Republik, Estland, Ungarn, Lettland, Litauen, Malta, Polen, Rumänien, die Slowakei und Slowenien [1]. (Hinweis: Die Republik Zypern hat juristisch Souveränität über die ganze Insel, da die Türkische Republik Nordzypern international nicht anerkannt wird.)

  4. In Vivo Monitoring Program Manual, PNL-MA-574, Rev 5.1

    SciTech Connect

    Lynch, Timothy P.

    2011-09-12

    The following sections provide an overview of the administration for the In Vivo Monitoring Program (IVMP) for Hanford. This includes the organizational structure and program responsibilities; coordination of in vivo measurements; scheduling measurements; performing measurements; reporting results; and quality assurance.

  5. Die kreiselnde Büroklammer: Spielwiese

    NASA Astrophysics Data System (ADS)

    Ucke, Christian; Schlichting, Hans-Joachim

    2005-01-01

    Mit Büroklammern lassen sich einfach und schnell physikalische Experimente realisieren. Vorgestellt werden zwei ungewöhnliche Kreisel (darunter ein Stehaufkreisel), die sich in wenigen Minuten biegen lassen. Weiterhin lässt sich mit Büroklammern die schon von Leibniz abgeleitete Kettenlinie simulieren. Mit etwas Mehraufwand lässt sich eine Hängebrücke bauen, bei der sich für das Tragkabel eine Parabel als Kurvenform ergibt. Im Internet sind Programme verfügbar, mit denen sich Kettenlinie und Hängeparabel simulieren lassen.

  6. Die casting die deflections: Prediction and attenuation. Final report, July 1, 1995--September 30, 1997

    SciTech Connect

    Miller, R.A.; Ahuett-Garza, H.; Choudhury, A.K.; Dedhia, S.

    1998-05-01

    This report summarizes two years of research intended to develop methods to model and predict the deflection patterns in die casting dies. No comprehensive analysis of this type had previously been completed. The die casting process is complex and involves numerous mechanical and thermal phenomena that effect the mechanical behavior of the die. A critical activity in this work was sorting out and evaluating the relative contributions of the various mechanisms to die deflections. This evaluation was accomplished through a series of simple engineering analyses based primarily on the order of magnitude of the influence of each load considered on die deflections. A modeling approach incorporating commercially available finite element analysis software was developed and tested. The model evolved by testing simple models against more comprehensive models and against the limited experimental data that is available. The development of the modeling approach lead to consideration of the die casting machine in more detail than was originally anticipated. The machine is critical and cannot be ignored. A simplified model described as a spring/platen model was developed to account for the machine platens, tie bars, and toggles. The characteristics of this model are described and predictions based on this model are compared against full machine models and measured deflections of machine platens. Details of the modeling approach and the various case studies are provided in the report and in several publications that have resulted from the work.

  7. Wissenschaft, die unsere Kultur verändert. Tiefenschichten des Streits um die Evolutionstheorie

    NASA Astrophysics Data System (ADS)

    Patzelt, Werner J.

    Die Evolutionstheorie ist eine der erfolgreichsten wissenschaftlichen Theorien. Sie erlaubt es, unsere Herkunft zu verstehen und riskante Merkmale gerade der menschlichen Spezies zu begreifen. Zugleich ist die Evolutionstheorie eine der umstrittensten Theorien. Das liegt nicht an ihrer empirischen Tragfähigkeit, sondern an ihrem Gegenstand. Sie handelt nämlich nicht nur - wie Hunderte andere wissenschaftliche Theorien - von der "Welt da draußen“, sondern vor allem auch von uns selbst und von unserem Platz in dieser Welt. Den einen gilt sie obendrein als Überwinderin religiösen Aberglaubens, den anderen als neuer Zugang zu Gott und seinem Wirken in der Welt. Ferner sehen die einen in der Evolution eine unbezweifelbare Tatsache gleich der Schwerkraft oder dem Holocaust, die anderen aber eine - noch oder dauerhaft - unbewiesene Hypothese oder gar eine falsche Schöpfungslehre. Und während die meisten Streitfragen solcher Art nach wechselseitig akzeptierten Regeln ‚normaler Wissenschaft‘ geklärt werden, wird bei der Frage nach dem Woher unserer Spezies und Kultur die intellektuelle Zuständigkeit von Wissenschaft mitunter überhaupt bezweifelt. Anscheinend geht es schon um recht tiefe Schichten unserer Kultur und nicht nur der wissenschaftlichen, wenn - wie seit 150 Jahren - um die Evolutionstheorie gestritten wird. Wie sehen diese Schichten aus?

  8. Effect of Die Strength and Work Piece Strength on the Wear of Hot Forging Dies

    NASA Astrophysics Data System (ADS)

    Levy, B. S.; Van Tyne, C. J.

    2015-01-01

    The effect of the strength ratio extracted from an Archard model for wear is used to describe the wear rates expected in hot forging dies. In the current study, the strength ratio is the strength of the hot forging die to the strength of the work piece. Three hot forging die steels are evaluated. The three die steels are FX, 2714, and WF. To determine the strength of the forging die, a continuous function has been developed that describes the yield strength of three die steels for temperatures from 600 to 700 °C and for times up to 20 h (i.e., tempering times of up to 20 h). The work piece material is assumed to be AISI 1045. Based on the analysis, the wear resistance of WF should be superior and FX should be slightly better than 2714. Decreasing the forging temperature increases the strength ratio, because the strength of the die surface increases faster than the flow strength of AISI 1045. The increase in the strength ratio indicates a decrease in the expected wear rate.

  9. Investigation of Die Stress Profiles during Powder Compaction using Instrumented Die

    SciTech Connect

    Hong, Sung-tae; Hovanski, Yuri; Lavender, Curt A.; Weil, K. Scott

    2008-06-01

    The radial stress profile in a cylindrical die during compaction of titanium (Ti) powder was investigated by experiments. The concept of an instrumented die was extended to design an enhanced instrumented die. Custom-made strain gage pins were used to measure the radial stress during powder compaction. The test fixture was designed to simulate double-action pressing. The measured die stress profile for Ti powder was compared with that for a commercially available iron (Fe) powder. The stress history shows that an appreciable residual stress remains in the die in the radial direction after the axial compaction stress is removed from the powder. Furthermore, the radial stress profile in the die, while under maximum axial compaction stress, is more uniform across the height of the Fe compact than that of the Ti compact. In addition, the residual stress profile in the die in the radial direction reduces symmetrically in both directions beyond the height of the compact for both powders. Finally, the Ti powder shows a significantly higher frictional coefficient at the maximum axial compaction stress, and consequently a higher maximum axial ejection stress than the Fe powder.

  10. In vivo endocrine effects of naphthenic acids in fish.

    PubMed

    Knag, Anne Christine; Sebire, Marion; Mayer, Ian; Meier, Sonnich; Renner, Patrick; Katsiadaki, Ioanna

    2013-11-01

    Oil pollution from various sources, including exploration, production and transportation, is a growing global concern. The highest toxicity of hydrocarbon pollutants is associated with the water-soluble phase compounds, including naphthenic acids, a known component found in all hydrocarbon deposits. Recently, naphthenic acids (NAs) have shown estrogenic and anti-androgenic effects in vitro. For this reason we investigated the potential effects of two commercial mixtures of naphthenic acids on fish in vivo, using the three-spined stickleback (Gasterosteus aculeatus) as a model species. Anti-androgenic and estrogenic properties of tested compounds were evaluated using the androgenized female stickleback screen (AFSS) and a variant of the 21-d fish screen (TG230) respectively. One-dimensional gas chromatography-mass spectrometry (GC-MS) showed that the complex commercial NAs mixtures were dominated by acyclic carboxylic acids. In one experiment (freshwater) we found a clear effect of NA exposure on spiggin levels; this was contrary to our hypothesis since NAs enhanced the androgenic potency of DHT (when co-administered) without inducing spiggin when tested in the absence of DHT. Exposure to NAs did not have a statistically significant effect on vitellogenin (Vtg) production in male stickleback, although the Vtg responses were increasing with increasing exposure concentrations. This study shows that in contrast to previous in vitro data, NAs did not exhibit either estrogenic or anti-androgenic properties in vivo, at the concentrations tested. On the contrary, at least in freshwater, NAs appear to have an overall androgenic effect that is not mediated via the androgen receptor involved in spiggin synthesis. Possible reasons for this discrepancy between in vitro and in vivo results as well as between our studies are discussed. PMID:24034895

  11. Villin Severing Activity Enhances Actin-based Motility In Vivo

    PubMed Central

    Revenu, Céline; Courtois, Matthieu; Michelot, Alphée; Sykes, Cécile; Louvard, Daniel

    2007-01-01

    Villin, an actin-binding protein associated with the actin bundles that support microvilli, bundles, caps, nucleates, and severs actin in a calcium-dependant manner in vitro. We hypothesized that the severing activity of villin is responsible for its reported role in enhancing cell plasticity and motility. To test this hypothesis, we chose a loss of function strategy and introduced mutations in villin based on sequence comparison with CapG. By pyrene-actin assays, we demonstrate that this mutant has a strongly reduced severing activity, whereas nucleation and capping remain unaffected. The bundling activity and the morphogenic effects of villin in cells are also preserved in this mutant. We thus succeeded in dissociating the severing from the three other activities of villin. The contribution of villin severing to actin dynamics is analyzed in vivo through the actin-based movement of the intracellular bacteria Shigella flexneri in cells expressing villin and its severing variant. The severing mutations abolish the gain of velocity induced by villin. To further analyze this effect, we reconstituted an in vitro actin-based bead movement in which the usual capping protein is replaced by either the wild type or the severing mutant of villin. Confirming the in vivo results, villin-severing activity enhances the velocity of beads by more than two-fold and reduces the density of actin in the comets. We propose a model in which, by severing actin filaments and capping their barbed ends, villin increases the concentration of actin monomers available for polymerization, a mechanism that might be paralleled in vivo when an enterocyte undergoes an epithelio-mesenchymal transition. PMID:17182858

  12. Nitroxoline induces apoptosis and slows glioma growth in vivo

    PubMed Central

    Lazovic, Jelena; Guo, Lea; Nakashima, Jonathan; Mirsadraei, Leili; Yong, William; Kim, Hyun J.; Ellingson, Benjamin; Wu, Hong; Pope, Whitney B.

    2015-01-01

    Background Nitroxoline is an FDA-approved antibiotic with potential antitumor activity. Here we evaluated whether nitroxoline has antiproliferative properties on glioma cell growth in vitro and in vivo using glioma cell lines and a genetically engineered PTEN/KRAS mouse glioma model. Methods The effect of nitroxoline treatment on U87 and/or U251 glioma cell proliferation, cell-cycle arrest, invasion, and ability to induce an apoptotic cascade was determined in vitro. Magnetic resonance imaging was used to measure glioma volumes in genetically engineered PTEN/KRAS mice prior to and after nitroxoline therapy. Induction of apoptosis by nitroxoline was evaluated at the end of treatment using terminal deoxyribonucleotidyl transferase (TDT)-mediated dUTP-digoxigenin nick end labeling (TUNEL). Results Nitroxoline inhibited the proliferation and invasion of glioblastoma cells in a time- and dose-dependent manner in vitro. Growth inhibition was associated with cell-cycle arrest in G1/G0 phase and induction of apoptosis via caspase 3 and cleaved poly(ADP-ribose) polymerase. In vivo, nitroxoline-treated mice had no increase in tumor volume after 14 days of treatment, whereas tumor volumes doubled in control mice. Histological examination revealed 15%–20% TUNEL-positive cells in nitroxoline-treated mice, compared with ∼5% in the control group. Conclusion Nitroxoline induces apoptosis and inhibits glioma growth in vivo and in vitro. As an already FDA-approved treatment for urinary tract infections with a known safety profile, nitroxoline could move quickly into clinical trials pending confirmatory studies. PMID:25074541

  13. Pharmacological Modulation of Hemodynamics in Adult Zebrafish In Vivo

    PubMed Central

    Brönnimann, Daniel; Dellenbach, Christian; Saveljic, Igor; Rieger, Michael; Rohr, Stephan; Filipovic, Nenad; Djonov, Valentin

    2016-01-01

    Introduction Hemodynamic parameters in zebrafish receive increasing attention because of their important role in cardiovascular processes such as atherosclerosis, hematopoiesis, sprouting and intussusceptive angiogenesis. To study underlying mechanisms, the precise modulation of parameters like blood flow velocity or shear stress is centrally important. Questions related to blood flow have been addressed in the past in either embryonic or ex vivo-zebrafish models but little information is available for adult animals. Here we describe a pharmacological approach to modulate cardiac and hemodynamic parameters in adult zebrafish in vivo. Materials and Methods Adult zebrafish were paralyzed and orally perfused with salt water. The drugs isoprenaline and sodium nitroprusside were directly applied with the perfusate, thus closely resembling the preferred method for drug delivery in zebrafish, namely within the water. Drug effects on the heart and on blood flow in the submental vein were studied using electrocardiograms, in vivo-microscopy and mathematical flow simulations. Results Under control conditions, heart rate, blood flow velocity and shear stress varied less than ± 5%. Maximal chronotropic effects of isoprenaline were achieved at a concentration of 50 μmol/L, where it increased the heart rate by 22.6 ± 1.3% (n = 4; p < 0.0001). Blood flow velocity and shear stress in the submental vein were not significantly increased. Sodium nitroprusside at 1 mmol/L did not alter the heart rate but increased blood flow velocity by 110.46 ± 19.64% (p = 0.01) and shear stress by 117.96 ± 23.65% (n = 9; p = 0.03). Discussion In this study, we demonstrate that cardiac and hemodynamic parameters in adult zebrafish can be efficiently modulated by isoprenaline and sodium nitroprusside. Together with the suitability of the zebrafish for in vivo-microscopy and genetic modifications, the methodology described permits studying biological processes that are dependent on hemodynamic

  14. In Vivo Ultrasonic Detection of Polyurea Crosslinked Silica Aerogel Implants

    PubMed Central

    Sabri, Firouzeh; Sebelik, Merry E.; Meacham, Ryan; Boughter, John D.; Challis, Mitchell J.; Leventis, Nicholas

    2013-01-01

    Background Polyurea crosslinked silica aerogels are highly porous, lightweight, and mechanically strong materials with great potential for in vivo applications. Recent in vivo and in vitro studies have demonstrated the biocompatibility of this type of aerogel. The highly porous nature of aerogels allows for exceptional thermal, electric, and acoustic insulating capabilities that can be taken advantage of for non-invasive external imaging techniques. Sound-based detection of implants is a low cost, non-invasive, portable, and rapid technique that is routinely used and readily available in major clinics and hospitals. Methodology In this study the first in vivo ultrasound response of polyurea crosslinked silica aerogel implants was investigated by means of a GE Medical Systems LogiQe diagnostic ultrasound machine with a linear array probe. Aerogel samples were inserted subcutaneously and sub-muscularly in a) fresh animal model and b) cadaveric human model for analysis. For comparison, samples of polydimethylsiloxane (PDMS) were also imaged under similar conditions as the aerogel samples. Conclusion/significance Polyurea crosslinked silica aerogel (X-Si aerogel) implants were easily identified when inserted in either of the regions in both fresh animal model and cadaveric model. The implant dimensions inferred from the images matched the actual size of the implants and no apparent damage was sustained by the X-Si aerogel implants as a result of the ultrasonic imaging process. The aerogel implants demonstrated hyperechoic behavior and significant posterior shadowing. Results obtained were compared with images acquired from the PDMS implants inserted at the same location. PMID:23799093

  15. Subacute Cardiotoxicity of Yessotoxin: In Vitro and in Vivo Studies.

    PubMed

    Ferreiro, Sara F; Vilariño, Natalia; Carrera, Cristina; Louzao, M Carmen; Cantalapiedra, Antonio G; Santamarina, Germán; Cifuentes, J Manuel; Vieira, Andrés C; Botana, Luis M

    2016-06-20

    Yessotoxin (YTX) is a marine phycotoxin produced by dinoflagellates and accumulated in filter feeding shellfish. Although no human intoxication episodes have been reported, YTX content in shellfish is regulated by many food safety authorities due to their worldwide distribution. YTXs have been related to ultrastructural heart damage in vivo, but the functional consequences in the long term have not been evaluated. In this study, we explored the accumulative cardiotoxic potential of YTX in vitro and in vivo. Preliminary in vitro evaluation of cardiotoxicity was based on the effect on hERG (human ether-a-go-go related gene) channel trafficking. In vivo experiments were performed in rats that received repeated administrations of YTX followed by recordings of electrocardiograms, arterial blood pressure, plasmatic cardiac biomarkers, and analysis of myocardium structure and ultrastructure. Our results showed that an exposure to 100 nM YTX for 12 or 24 h caused an increase of extracellular surface hERG channels. Furthermore, remarkable bradycardia and hypotension, structural heart alterations, and increased plasma levels of tissue inhibitor of metalloproteinases-1 were observed in rats after four intraperitoneal injections of YTX at doses of 50 or 70 μg/kg that were administered every 4 days along a period of 15 days. Therefore, and for the first time, YTX-induced subacute cardiotoxicity is supported by evidence of cardiovascular function alterations related to its repeated administration. Considering international criteria for marine toxin risk estimation and that the regulatory limit for YTX has been recently raised in many countries, YTX cardiotoxicity might pose a health risk to humans and especially to people with previous cardiovascular risk. PMID:27104637

  16. In vivo characterization of regenerative peripheral nerve interface function

    NASA Astrophysics Data System (ADS)

    Ursu, Daniel C.; Urbanchek, Melanie G.; Nedic, Andrej; Cederna, Paul S.; Gillespie, R. Brent

    2016-04-01

    Objective. Regenerative peripheral nerve interfaces (RPNIs) are neurotized free autologous muscle grafts equipped with electrodes to record myoelectric signals for prosthesis control. Viability of rat RPNI constructs have been demonstrated using evoked responses. In vivo RPNI characterization is the next critical step for assessment as a control modality for prosthetic devices. Approach. Two RPNIs were created in each of two rats by grafting portions of free muscle to the ends of divided peripheral nerves (peroneal in the left and tibial in the right hind limb) and placing bipolar electrodes on the graft surface. After four months, we examined in vivo electromyographic signal activity and compared these signals to muscular electromyographic signals recorded from autologous muscles in two rats serving as controls. An additional group of two rats in which the autologous muscles were denervated served to quantify cross-talk in the electrode recordings. Recordings were made while rats walked on a treadmill and a motion capture system tracked the hind limbs. Amplitude and periodicity of signals relative to gait were quantified, correlation between electromyographic and motion recording were assessed, and a decoder was trained to predict joint motion. Main Results. Raw RPNI signals were active during walking, with amplitudes of 1 mVPP, and quiet during standing, with amplitudes less than 0.1 mVPP. RPNI signals were periodic and entrained with gait. A decoder predicted bilateral ankle motion with greater than 80% reliability. Control group signal activity agreed with literature. Denervated group signals remained quiescent throughout all evaluations. Significance. In vivo myoelectric RPNI activity encodes neural activation patterns associated with gait. Signal contamination from muscles adjacent to the RPNI is minimal, as demonstrated by the low amplitude signals obtained from the Denervated group. The periodicity and entrainment to gait of RPNI recordings suggests the

  17. The sustained phase of tyrosine hydroxylase activation in vivo.

    PubMed

    Ong, Lin Kooi; Sominsky, Luba; Dickson, Phillip W; Hodgson, Deborah M; Dunkley, Peter R

    2012-09-01

    Tyrosine hydroxylase (TH) is the rate-limiting enzyme in the biosynthetic pathway for catecholamine synthesis. Stress triggers an increase in TH activity, resulting in increased release of catecholamines from both neurons and the adrenal medulla. In response to stress three phases of TH activation have been identified (acute, sustained and chronic) and each phase has a unique mechanism. The acute and chronic phases have been studied in vivo in a number of animal models, but to date the sustained phase has only been characterised in vitro. We aimed to investigate the effects of dual exposure to lipopolysaccharide (LPS) in neonatal rats on TH protein, TH phosphorylation at serine residues 19, 31 and 40 and TH activity in the adrenal gland over the sustained phase. Wistar rats were administered LPS (0.05 mg/kg, intraperitoneal injection) or an equivolume of non-pyrogenic saline on days 3 and 5 postpartum. Adrenal glands were collected at 4, 24 and 48 h after the drug exposure on day 5. Neonatal LPS treatment resulted in increases in TH phosphorylation of Ser40 at 4 and 24 h, TH phosphorylation of Ser31 at 24 h, TH activity at 4 and 24 h and TH protein at 48 h. We therefore have provided evidence for the first time that TH phosphorylation at Ser31 and Ser40 occurs for up to 24 h in vivo and leads to TH activation independent of TH protein synthesis, suggesting that the sustained phase of TH activation occurs in vivo. PMID:22684282

  18. In vivo cellular imaging with microscopes enabled by MEMS scanners

    NASA Astrophysics Data System (ADS)

    Ra, Hyejun

    High-resolution optical imaging plays an important role in medical diagnosis and biomedical research. Confocal microscopy is a widely used imaging method for obtaining cellular and sub-cellular images of biological tissue in reflectance and fluorescence modes. Its characteristic optical sectioning capability also enables three-dimensional (3-D) image reconstruction. However, its use has mostly been limited to excised tissues due to the requirement of high numerical aperture (NA) lenses for cellular resolution. Microscope miniaturization can enable in vivo imaging to make possible early cancer diagnosis and biological studies in the innate environment. In this dissertation, microscope miniaturization for in vivo cellular imaging is presented. The dual-axes confocal (DAC) architecture overcomes limitations of the conventional single-axis confocal (SAC) architecture to allow for miniaturization with high resolution. A microelectromechanical systems (MEMS) scanner is the central imaging component that is key in miniaturization of the DAC architecture. The design, fabrication, and characterization of the two-dimensional (2-D) MEMS scanner are presented. The gimbaled MEMS scanner is fabricated on a double silicon-on-insulator (SOI) wafer and is actuated by self-aligned vertical electrostatic combdrives. The imaging performance of the MEMS scanner in a DAC configuration is shown in a breadboard microscope setup, where reflectance and fluorescence imaging is demonstrated. Then, the MEMS scanner is integrated into a miniature DAC microscope. The whole imaging system is integrated into a portable unit for research in small animal models of human biology and disease. In vivo 3-D imaging is demonstrated on mouse skin models showing gene transfer and siRNA silencing. The siRNA silencing process is sequentially imaged in one mouse over time.

  19. In Vivo Characterization of a Vertebrate Ultra-conserved Enhancer

    SciTech Connect

    Poulin, Francis; Nobrega, Marcelo A.; Plajzer-Frick, Ingrid; Holt, Amy; Afzal, Veena; Rubin, Edward M.; Pennacchio, Len

    2004-10-01

    Genomic sequence comparisons between human, mouse and pufferfish (Takifugu rubripes (Fugu))have revealed a set of extremely conserved noncoding sequences. While this high degree of sequence conservation suggests severe evolutionary constraint and predicts a lack of tolerance to change in order to retain in vivo functionality, such elements have been minimally explored experimentally. In this study, we describe the in-depth characterization of an ancient conserved enhancer, Dc2 located near the dachshund gene, which displays a human-Fugu identity of 84 percent over 424 basepairs (bp). In addition to this large overall conservation, we find that Dc2 is characterized by the presence of a large block of sequence (144 bp) that is completely identical between human, mouse, chicken, zebrafish and Fugu. Through the testing of reporter vector constructs in transgenic mice, we observed that the 424 bp Dc2 conserved element is necessary and sufficient for brain tissue enhancer activity. In vivo analyses also revealed that the 144 bp 100 percent conserved sequence is necessary, but not sufficient, to replicate Dc2 enhancer function. However, the introduction of two separate 16 bp insertions into the highly conserved enhancer core did not cause any detectable modification of its in vivo activity. Our observations indicate that the 144 bp 100 percent conserved element is tolerant of change at least at the resolution of this transgenic mouse assay and suggest that purifying selection on Dc2 sequence might not be as strong as we predicted or that some unknown property also constrains this highly conserved enhancer sequence.

  20. In Vivo Evaluation of Wearable Head Impact Sensors.

    PubMed

    Wu, Lyndia C; Nangia, Vaibhav; Bui, Kevin; Hammoor, Bradley; Kurt, Mehmet; Hernandez, Fidel; Kuo, Calvin; Camarillo, David B

    2016-04-01

    Inertial sensors are commonly used to measure human head motion. Some sensors have been tested with dummy or cadaver experiments with mixed results, and methods to evaluate sensors in vivo are lacking. Here we present an in vivo method using high speed video to test teeth-mounted (mouthguard), soft tissue-mounted (skin patch), and headgear-mounted (skull cap) sensors during 6-13 g sagittal soccer head impacts. Sensor coupling to the skull was quantified by displacement from an ear-canal reference. Mouthguard displacements were within video measurement error (<1 mm), while the skin patch and skull cap displaced up to 4 and 13 mm from the ear-canal reference, respectively. We used the mouthguard, which had the least displacement from skull, as the reference to assess 6-degree-of-freedom skin patch and skull cap measurements. Linear and rotational acceleration magnitudes were over-predicted by both the skin patch (with 120% NRMS error for [Formula: see text], 290% for [Formula: see text]) and the skull cap (320% NRMS error for [Formula: see text], 500% for [Formula: see text]). Such over-predictions were largely due to out-of-plane motion. To model sensor error, we found that in-plane skin patch linear acceleration in the anterior-posterior direction could be modeled by an underdamped viscoelastic system. In summary, the mouthguard showed tighter skull coupling than the other sensor mounting approaches. Furthermore, the in vivo methods presented are valuable for investigating skull acceleration sensor technologies. PMID:26289941

  1. Effects of survivin on angiogenesis in vivo and in vitro

    PubMed Central

    Li, Zhui; Ren, Wei; Zeng, Qiu; Chen, Siyu; Zhang, Mao; Zhao, Yu; Cheng, Jun; Wang, Xuehu

    2016-01-01

    Objective: In this study, the effects of survivin (SVV) on angiogenesis were evaluated in vitro and in vivo. Methods: The adenovirus (Ad)-mediated murine SVV gene was transfected into rat aortic endothelial cells (RAECs). RAECs expressing green fluorescent protein after transfection with Ad served as a negative control and those without transfection as a blank control. Then, the SVV mRNA was detected by quantitative real time RT-PCR. The SVV protein, cell cycle and apoptosis related proteins, and matrix metalloproteinase (MMPs) were detected by western blot assay. Immunofluorescence staining was conducted for proliferating cell nuclear antigen and MTT assay for cell viability. Transwell and matrigel chamber assay were employed to assess the migration and invasion of cells after transfection. TUNEL staining and flow cytometry were performed to detect the apoptotic REACs after treatment with anti-Fas antibody. Tube formation in matrigel membranes and matrigel plugs assay in nude mice were employed to confirm the angiogenic capacity in vitro and in vivo, respectively. Results: The mRNA and protein expressions of SVV increased significantly in SVV transfected cells. The SVV transfected cells showed increased cell proliferation, up-regulated expressions of cell cycle proteins, enhanced invasiveness and migration activities and increased expressions of MMP-2, 7 and 9. In addition, SVV protected against apoptosis of RAECs by inactivating caspase-3, 8 and 9. The tube formation and matrigel plugs assays showed SVV significantly increased blood vessels in vitro and in vivo. Conclusion: SVV may act as an angiogenic factor and used for therapeutic angiogenesis in peripheral arterial diseases. PMID:27158325

  2. Novel in vivo techniques to visualize kidney anatomy and function.

    PubMed

    Peti-Peterdi, János; Kidokoro, Kengo; Riquier-Brison, Anne

    2015-07-01

    Intravital imaging using multiphoton microscopy (MPM) has become an increasingly popular and widely used experimental technique in kidney research over the past few years. MPM allows deep optical sectioning of the intact, living kidney tissue with submicron resolution, which is unparalleled among intravital imaging approaches. MPM has solved a long-standing critical technical barrier in renal research to study several complex and inaccessible cell types and anatomical structures in vivo in their native environment. Comprehensive and quantitative kidney structure and function MPM studies helped our better understanding of the cellular and molecular mechanisms of the healthy and diseased kidney. This review summarizes recent in vivo MPM studies with a focus on the glomerulus and the filtration barrier, although select, glomerulus-related renal vascular and tubular functions are also mentioned. The latest applications of serial MPM of the same glomerulus in vivo, in the intact kidney over several days, during the progression of glomerular disease are discussed. This visual approach, in combination with genetically encoded fluorescent markers of cell lineage, has helped track the fate and function (e.g., cell calcium changes) of single podocytes during the development of glomerular pathologies, and provided visual proof for the highly dynamic, rather than static, nature of the glomerular environment. Future intravital imaging applications have the promise to further push the limits of optical microscopy, and to advance our understanding of the mechanisms of kidney injury. Also, MPM will help to study new mechanisms of tissue repair and regeneration, a cutting-edge area of kidney research. PMID:25738253

  3. In vivo imaging of tumor vascular endothelial cells

    NASA Astrophysics Data System (ADS)

    Zhao, Dawen; Stafford, Jason H.; Zhou, Heling; Thorpe, Philip E.

    2013-02-01

    Phosphatidylserine (PS), normally restricted to the inner leaflet of the plasma membrane, becomes exposed on the outer surface of viable (non-apoptotic) endothelial cells in tumor blood vessels, probably in response to oxidative stresses present in the tumor microenvironment. In the present study, we optically imaged exposed PS on tumor vasculature in vivo using PGN635, a novel human monoclonal antibody that targets PS. PGN635 F(ab')2 was labeled with the near infrared (NIR) dye, IRDye 800CW. Human glioma U87 cells or breast cancer MDA-MB-231 cells were implanted subcutaneously or orthotopically into nude mice. When the tumors reached ~5 mm in diameter, 800CW- PGN635 was injected via a tail vein and in vivo dynamic NIR imaging was performed. For U87 gliomas, NIR imaging allowed clear detection of tumors as early as 4 h later, which improved over time to give a maximal tumor/normal ratio (TNR = 2.9 +/- 0.5) 24 h later. Similar results were observed for orthotopic MDA-MB-231 breast tumors. Localization of 800CW-PGN635 to tumors was antigen specific since 800CW-Aurexis, a control probe of irrelevant specificity, did not localize to the tumors, and pre-administration of unlabeled PGN635 blocked the uptake of 800CW-PGN635. Fluorescence microscopy confirmed that 800CW-PGN635 was binding to PS-positive tumor vascular endothelium. Our studies suggest that tumor vasculature can be successfully imaged in vivo to provide sensitive tumor detection.

  4. In vivo porcine training model for cranial neurosurgery.

    PubMed

    Regelsberger, Jan; Eicker, Sven; Siasios, Ioannis; Hänggi, Daniel; Kirsch, Matthias; Horn, Peter; Winkler, Peter; Signoretti, Stefano; Fountas, Kostas; Dufour, Henry; Barcia, Juan A; Sakowitz, Oliver; Westermaier, Thomas; Sabel, Michael; Heese, Oliver

    2015-01-01

    Supplemental education is desirable for neurosurgical training, and the use of human cadaver specimen and virtual reality models is routine. An in vivo porcine training model for cranial neurosurgery was introduced in 2005, and our recent experience with this unique model is outlined here. For the first time, porcine anatomy is illustrated with particular respect to neurosurgical procedures. The pros and cons of this model are described. The aim of the course was to set up a laboratory scenery imitating an almost realistic operating room in which anatomy of the brain and neurosurgical techniques in a mentored environment free from time constraints could be trained. Learning objectives of the course were to learn about the microsurgical techniques in cranial neurosurgery and the management of complications. Participants were asked to evaluate the quality and utility of the programme via standardized questionnaires by a grading scale from A (best) to E (worst). In total, 154 residents have been trained on the porcine model to date. None of the participants regarded his own residency programme as structured. The bleeding and complication management (97%), the realistic laboratory set-up (89%) and the working environment (94%) were favoured by the vast majority of trainees and confirmed our previous findings. After finishing the course, the participants graded that their skills in bone drilling, dissecting the brain and preserving cerebral vessels under microscopic magnification had improved to level A and B. In vivo hands-on courses, fully equipped with microsurgical instruments, offer an outstanding training opportunity in which bleeding management on a pulsating, vital brain represents a unique training approach. Our results have shown that education programmes still lack practical training facilities in which in vivo models may act as a complementary approach in surgical training. PMID:25240530

  5. Heme oxygenase metabolites inhibit tubuloglomerular feedback in vivo.

    PubMed

    Wang, Hong; Garvin, Jeffrey L; D'Ambrosio, Martin A; Falck, John R; Leung, Pablo; Liu, Ruisheng; Ren, YiLin; Carretero, Oscar A

    2011-04-01

    Tubuloglomerular feedback (TGF) is a renal autoregulatory mechanism that constricts the afferent arteriole in response to increases in distal NaCl. Heme oxygenases (HO-1 and HO-2) release carbon monoxide (CO) and biliverdin, which may help control renal function. We showed in vitro that HO products inhibit TGF; however, we do not know whether this also occurs in vivo or the mechanism(s) involved. We hypothesized that in vivo HO-1 and HO-2 in the nephron inhibit TGF via release of CO and biliverdin. We first performed laser capture microdissection followed by real-time PCR and found that both HO-1 and HO-2 are expressed in the macula densa. We next performed micropuncture experiments in vivo on individual rat nephrons, adding different compounds to the perfusate, and found that an HO inhibitor, stannous mesoporphyrin (SnMP), potentiated TGF (P < 0.05, SnMP vs. control). The CO-releasing molecule (CORM)-3 partially inhibited TGF at 50 μmol/l (P < 0.01, CORM-3 vs. control) and blocked it completely at higher doses. A soluble guanylyl cyclase (sGC) inhibitor, LY83583, blocked the inhibitory effect of CORM-3 on TGF. Biliverdin also partially inhibited TGF (P < 0.01, biliverdin vs. control), most likely attributable to decreased superoxide (O(2)(-)) because biliverdin was rendered ineffective by tempol, a O(2)(-) dismutase mimetic. We concluded that HO-1 and HO-2 in the nephron inhibit TGF by releasing CO and biliverdin. The inhibitory effect of CO on TGF is mediated by the sGC/cGMP signaling pathway, whereas biliverdin probably acts by reducing O(2)(-). PMID:21239629

  6. Associations between in-vivo glenohumeral joint motion and morphology.

    PubMed

    Peltz, Cathryn D; Divine, George; Drake, Anne; Ramo, Nicole L; Zauel, Roger; Moutzouros, Vasilios; Bey, Michael J

    2015-09-18

    Joint morphology has a significant influence on joint motion and may contribute to the development of rotator cuff pathology, but the relationships between glenohumeral joint (GHJ) morphology and in-vivo GHJ motion are not well understood. The objectives of this study were to assess measures of joint morphology and their relationship with in-vivo joint motion in two populations: shoulders with intact rotator cuffs (n=48) and shoulders with rotator cuff pathology (n=36, including 5 symptomatic tears, 9 asymptomatic tears and 22 repaired tears). GHJ morphology was measured from CT-based three-dimensional models of the humerus and scapula. In-vivo GHJ motion was measured during shoulder abduction using biplane x-ray imaging. Associations between GHJ morphology and motion were assessed with univariate and best subsets regression. The only morphological difference identified between the populations was the critical shoulder angle (intact: 34.5 ± 4.7°, pathologic: 36.9 ± 5.0°, p=0.03), which is consistent with previous research. In intact shoulders, the superior/inferior (S/I) position of the humerus on the glenoid during shoulder abduction was significantly associated with the glenoid's S/I radius of curvature (p<0.01), conformity index (p<0.01), and stability angle (p<0.01). Furthermore, the S/I position of the humerus on the glenoid was negatively associated with the critical shoulder angle (p=0.04), which contradicts previous research. No significant associations between GHJ morphology and GHJ motion were detected in shoulders with rotator cuff tears. It is unknown if rotator cuff pathology compromises the relationships between GHJ morphology and motion, or if the absence of this relationship is a pre-existing condition that increases the likelihood of pathology. PMID:26189094

  7. Gastric ulcer localization: Potential use of in vivo labeling

    SciTech Connect

    Pera, A.; Rose, H.; Seavers, R.; Bekerman, C.; Pinsky, S.

    1984-01-01

    A previous work suggests that sucralfate labeled by binding to Tc-99m HSA permits the visualization of gastric ulcers. Potential problems with this technique are: 1) decreased binding of sucralfate to ulcer sites due to the labeling method of binding to exogenous protein (HSA); 2) overlying activity that may obscure identification of the ulcer. Because of these problems we have examined the possibility of direct in vivo Tc-99m labeling of sucralfate after it has already bound to the ulcer. In vitro studies were done to determine the binding of Tc-99m pertechnetate to sucralfate in the presence of tin in HCl solution at pHs comparable to those found in the stomach. Rapid and efficient labeling was achieved with 75-95% of the label bound to sucralfate at 30 minutes. In vivo studies were performed in rabbits with aspirin induced ulcers and in ulcer free human volunteers. The animal studies confirm that orally administered Tc-99m pertechnetate will bind to previously ingested sucralfate and that the labeled material will bind to the ulcers. Tc-99m pertechnetate was also shown to bind well to previously ingested sucralfate in humans. The results suggest that it is possible to label sucralfate in vivo. This method would offer the following advantages: 1) a simpler labeling procedure; 2) the potential of increased sensitivity by delaying the labeling until much of the sucralfate not bound to ulcer has passed, and thus decreasing the activity that remains in the stomach; and also by leaving the protein binding sites of the sucralfate free to interact with the ulcer since no exogenous protein is involved in labeling.

  8. Multimodal system for in vivo tumor imaging in mice

    NASA Astrophysics Data System (ADS)

    Autiero, Maddalena; Celentano, Luigi; Cozzolino, Rosanna; Laccetti, Paolo; Marotta, Marcello; Mettivier, Giovanni; Montesi, Maria Cristina; Riccio, Patrizia; Roberti, Giuseppe; Russo, Paolo

    2006-04-01

    We devised a multimodal planar imaging system for in vivo mouse imaging, employing four modalities: optical imaging, green and red fluorescence reflectance imaging, radionuclide imaging and X-ray radiography. We are testing separately, and then in a combined way, each detection mode, via in vivo mouse imaging, with the final purpose of identifying small implanted tumor masses, of providing early tumor detection and following metastatic dissemination. We describe the multimodal system and summarize its main performance, as assessed in our research work in the various stages of the development, in fluorescence and radionuclide tests on healthy or tumor bearing mice. For gamma-ray detection we used a semiconductor pixel detector (Medipix1 or Medipix2) that works in single photon counting. Laser-induced fluorescence reflectance imaging was performed in vivo using a pulsed light source to excite the fluorescence emission of injected hematoporphyrin (HP) compound, a CCD camera, a low pass filter and a commercial image analysis system. The bimodal system was used for the acquisition of combined images of the tumor area (fluorescence: animal top view; radionuclide: bottom view). It was shown that the tumor area can be imaged in a few minutes, with a few millimeter resolution (1 mm pinhole diameter), radioactively ( 99mTc radiotracer), and with the fluorescence system and that, in one case, only one of the two modalities is able to recognize the tumor. A phantom study for thyroid imaging with 125I source embedded in a simulated tissue indicated a spatial resolution of 1.25 mm FWHM with a 1 mm pinhole.

  9. Endothelial tubes assemble from intracellular vacuoles in vivo.

    PubMed

    Kamei, Makoto; Saunders, W Brian; Bayless, Kayla J; Dye, Louis; Davis, George E; Weinstein, Brant M

    2006-07-27

    The formation of epithelial tubes is crucial for the proper development of many different tissues and organs, and occurs by means of a variety of different mechanisms. Morphogenesis of seamless, properly patterned endothelial tubes is essential for the development of a functional vertebrate circulatory system, but the mechanism of vascular lumenization in vivo remains unclear. Evidence dating back more than 100 years has hinted at an important function for endothelial vacuoles in lumen formation. More than 25 years ago, in some of the first endothelial cell culture experiments in vitro, Folkman and Haudenschild described "longitudinal vacuoles" that "appeared to be extruded and connected from one cell to the next", observations confirmed and extended by later studies in vitro showing that intracellular vacuoles arise from integrin-dependent and cdc42/Rac1-dependent pinocytic events downstream of integrin-extracellular-matrix signalling interactions. Despite compelling data supporting a model for the assembly of endothelial tubes in vitro through the formation and fusion of vacuoles, conclusive evidence in vivo has been lacking, primarily because of difficulties associated with imaging the dynamics of subcellular endothelial vacuoles deep within living animals. Here we use high-resolution time-lapse two-photon imaging of transgenic zebrafish to examine how endothelial tubes assemble in vivo, comparing our results with time-lapse imaging of human endothelial-cell tube formation in three-dimensional collagen matrices in vitro. Our results provide strong support for a model in which the formation and intracellular and intercellular fusion of endothelial vacuoles drives vascular lumen formation. PMID:16799567

  10. In vivo target of benzylpenicillin in Gaffkya homari

    SciTech Connect

    Wrezel, P.W.

    1986-01-01

    It has been established that the DD-carboxypeptidase is the primary in vitro target of benzylpenicillin in Gaffkya homari. To determine whether this enzyme is also the primary target of benzylpenicillin in vivo, the effects of this ..beta..-lactam, cefmenoxime, cephalothin, and cefoxitin on growth were compared with their acylation of penicillin-binding protein (PBP) 9, the DD-carboxypeptidase. Results from experiments with membrane-walls indicated that PBP 9 is this enzyme and that it is the primary in vitro target of these ..beta..-lactams in the synthesis of SDS-insoluble peptidoglycan (PG). When intact cells were treated with benzylpenicillin, the MGIC (minimum growth inhibitor concentration) correlated with the B/sub 50/ values (concentration of (/sup 35/S)benzylpenicillin required to acylate a PBP by 50%) for PBPs 6 and 9. When intact cells were treated with cefmenoxime, cephalothin, or cefoxitin, the MGIC values correlated with the ED/sub 50/ values (concentration of unlabeled ..beta..-lactam required to reduce the subsequent binding of (/sup 35/S)benzylpenicillin by 50%) for PBP 6. In contrast, the MGIC values of these ..beta..-lactams did not correlate with the ED/sub 50/ values for PBP 9. It is suggested that (1) PBP 6 may be a primary target of growth inhibition by benzylpenicillin, cefmenoxime, cephalothin, and cefoxitin, that (2) PBP 9, the DD-carboxypeptidase is dispensable for growth under laboratory conditions, and that (3) PBP 9 does not appear to be a primary in vivo target of these ..beta..-lactams. PBP 6 appears to play a role in transpeptidation in vivo because cross-linking and wall PG synthesis occur at a concentration of cefoxitin that acylates all PBPs except PBP 6.

  11. Breast in vivo dosimetry by a portal ionization chamber

    SciTech Connect

    Grimaldi, Luca; D'Onofrio, Guido; Cilla, Savino; Fidanzio, Andrea; Stimato, Gerardina; Azario, Luigi; Deodato, Francesco; Macchia, Gabriella; Morganti, Alessio; Piermattei, Angelo

    2007-03-15

    This work reports a practical method for the determination of the in vivo breast middle dose value, D{sub m}, on the beam central axis, using a signal S{sub t}, obtained by a small thimble ion chamber positioned at the center of the electronic portal imaging device, and irradiated by the x-ray beam transmitted through the patient. The use of a stable ion chamber reduces many of the disadvantages associated with the use of diodes as their periodic recalibration and positioning is time consuming. The method makes use of a set of correlation functions obtained by the ratios S{sub t}/D{sub m}, determined by irradiating cylindrical water phantoms with different diameters. The method proposed here is based on the determination of the water-equivalent thickness of the patient, along the beam central axis, by the treatment planning system that makes use of the electron densities obtained by a computed tomography scanner. The method has been applied for the breast in vivo dosimetry of ten patients treated with a manual intensity modulation with four asymmetric beams. In particular, two tangential rectangular fields were first delivered, thereafter a fraction of the dose (typically less than 10%) was delivered with two multi leaf-shaped beams which included only the mammarian tissue. Only the two rectangular fields were tested and for every checked field five measurements were carried out. Applying a continuous quality assurance program based on the tests of patient setup, machine settings and dose planning, the proposed method is able to verify agreements between the computed dose D{sub m,TPS} and the in vivo dose value D{sub m}, within 4%.

  12. Noninvasive laser coagulation of the canine vas deferens, in vivo

    NASA Astrophysics Data System (ADS)

    Cilip, Christopher M.; Ross, Ashley E.; Jarow, Jonathan P.; Fried, Nathaniel M.

    2010-02-01

    Development of a noninvasive vasectomy technique may eliminate male fear of complications (incision, bleeding, infection, and scrotal pain) and result in a more popular procedure. This study builds upon previously reported ex vivo tissue studies by exploring acute and short-term chronic in vivo canine studies. Isolation of the canine vas was achieved using a conventional vas ring clamp method. No perforation of the scrotal skin was necessary to occlude the vas. Laser radiation with a wavelength of 1075 nm, average power of 11.2 W, 500-ms pulse duration, 0.5 Hz pulse rate, and 3-mm-diameter spot was synchronized with cryogen spray cooling of the scrotal skin surface in a total of 8 dogs (n = 16 vasa) for a treatment time of 60 s. Burst pressure measurements were conducted at Days 0 and 21 (n = 8 vasa each day) to quantify the strength of vas closure. The vas was successfully thermally occluded in 15/16 (94%) procedures with 14/15 (93%) vas recording burst pressures above ejaculation pressure. One vas was not present, and another vas recorded a bursting pressure below ejaculation pressure. The coagulated vas bursting pressure averaged 283 +/- 34 mm Hg at Day 0 and 260 +/- 77 mm Hg at Day 21, significantly higher than reported vas ejaculation pressures of 136 +/- 29 mm Hg. Minor scrotal skin burns were observed during the recovery period. Noninvasive thermal occlusion of the vas is feasible in an in vivo canine model. Elimination of minor skin burns and longer term chronic in vivo canine studies are needed to confirm azospermia after vas occlusion without recanalization.

  13. Complexities of glucuronidation affecting in vitro in vivo extrapolation.

    PubMed

    Lin, Jiunn H; Wong, Bradley K

    2002-12-01

    Glucuronidation is responsible for the clearance of a diverse range of drug and chemicals whose topology confers properties that complicate in vitro-in vivo clearance correlations as compared to those possible for oxidative metabolism. The active site of the UGTs faces the inside of the luminal space of the endoplasmic reticulum, thus presenting diffusional barriers for substrates, the cosubstrate, UDPGA, and resultant glucuronide products. Transport processes for the cosubstrate UDPGA and glucuronidated products likely contribute to the well-known latency phenomena in which exogenous detergents or alamethicin are required for maximal UGT activity in microsomes. This complicates the extrapolation of results of in vitro clearance studies to the in vivo situation. Even with activation, the microsomal-based clearance values still underestimate the actual in vivo UGT-mediated clearance; therefore latency is not the only explanation for the poor correlation. Recent data indicate that hepatocytes are a promising in vitro system that can be used for the early evaluation of human clearance behavior of drug candidates. Both induction and inhibition of UGT-mediated clearance are a source of clinical drug-drug interactions. Emerging evidence indicates that the same mechanisms identified in the regulation of CYP enzymes also are involved in regulation of the UGTs, i.e., CAR, AH and probably PXR mediate regulation of UGT1A1, 1A6 and UGT2B7, respectively. In contrast to CYP-mediated interactions, with a few exceptions, the magnitude of UGT-mediated interactions are less than 2-fold because of the relatively high UGT Km values and substrate overlap among the multiple isozymes. PMID:12369890

  14. Dying other, dying self: creating culture and meaning in palliative healthcare.

    PubMed

    McCann, Christopher J; Adames, Hector Y

    2013-08-01

    Dying is an act of creativity, and we each die as cultural beings. Culture helps us create the meaning death requests of us. However, the dominant culture of the healthcare system views death as a failure of modern medicine, an event of unspeakable terror and taboo. Palliative clinicians must honor each dying person's cultural identity (as well as the person's family), not subject it to the dominant discourse of Western medicine. This article offers practical guidelines for palliative clinicians to do so, as well as a case vignette. PMID:22854052

  15. Continuing Education on Dying and Death.

    ERIC Educational Resources Information Center

    Chodil, Judith J.; Dulaney, Peggy E.

    1984-01-01

    "Dying and Death in Critical Care Practice" was a one-day continuing education offering designed for registered nurses who practiced in settings such as emergency rooms, intensive care units, coronary care units, and operating rooms. The workshop was part of a continuing education curriculum in critical care nursing. (SSH)

  16. Physiknobelei Kann man die Lichtausbreitung sehen?

    NASA Astrophysics Data System (ADS)

    Schlichting, H.-Joachim

    2003-07-01

    Des Menschen Sinne sind trügerisch. Das wussten schon die Philosophen der Antike, denen physikalische Zusammenhänge noch fremd waren. Doch auch in der heutigen aufgeklärten Zeit ereignen sich noch Dinge, bei denen man seinen Augen nicht traut.

  17. Dying in the age of choice.

    PubMed

    Black, Kathy; Csikai, Ellen L

    2015-01-01

    Due to the unprecedented increase in the United States aging demographics, many more people are living longer and reaching older ages than ever before. However, a longer life is not necessarily a better life, as the vast majority will face a period of prolonged deteriorating health prior to death. Although notable efforts have been underway that are designed to improve the end-of-life experience, increasing numbers of individuals express a desire and/or act upon an intent to end their lives precipitously. Though still limited, the options to actively participate in their own deaths are growing. Requests for a hastened death can occur among people of all ages and includes those with advanced illness as well as others wanting to die due to unbearable suffering. This article provides an overview of the ongoing discourse about the experience of dying faced by many older adults, including aspects frequently associated with "a good death." The limitations of established practices which seek to provide a "better" dying experience are identified followed by discussion of the growing availability of alternative options. Reflective considerations are presented to guide practice vis-à-vis the changing landscape surrounding options in dying. PMID:25869146

  18. Care of the Dying: A Swedish Perspective

    ERIC Educational Resources Information Center

    Feigenberg, Loma; Fulton, Robert

    1977-01-01

    This article illustrates various aspects of terminal care, and shows that rules and norms for such care do not exist today. The authors advocate the formulation of an aim for humane treatment of dying patients, and its application in a manner appropriate to Swedish medical concepts and Swedish conditions. (Author)

  19. Flexible, Ultra-Thin, Embedded Die Packaging

    NASA Astrophysics Data System (ADS)

    McPherson, Ryan J.

    As thin, flexible electronics solutions become more robust, their integration into everyday life becomes more likely. With possible applications in wearable electronics, biomedical sensors, or 'peel and stick' sensors, the reliability of these ultra-thin packages becomes paramount. Likewise, the density achievable with stacked packages benefits greatly from thinner die stacks. To this end, techniques previously developed have demonstrated packages with die thinned to approximately 20mum. Covered in this work are methods for thinning and packaging silicon die, as well as information on common materials used in these processes. The author's contribution is a fabrication process for embedding ultra-thin (approximately 10mum) silicon die in polyimide substrates. This method is fully illustrated in Chapter 3 and enumerated in the Appendix as a quick reference. Additionally, thermal cycle testing of passive daisy chain assemblies has shown promising reliability data. Packages were mounted in three alignments: flat, concave, and convex, and placed into thermal shock testing. Finally, the author discusses possible applications for this fabrication process, including the fabrication of multi-chip-modules.

  20. Asymmetric Die Grows Purer Silicon Ribbon

    NASA Technical Reports Server (NTRS)

    Kalejs, J. P.; Chalmers, B.; Surek, T.

    1983-01-01

    Concentration of carbide impurities in silicon ribbon is reduced by growing crystalline ribbon with die one wall higher than other. Height difference controls shape of meniscus at liquid/crystal interface and concentrates silicon carbide impurity near one of broad faces. Opposite face is left with above-average purity. Significantly improves efficiency of solar cells made from ribbon.